Astaxanthin Abstract Book 2013 - Cyanotech

The Medical Research of 

Astaxanthin 

© Copyright 2013 Cyanotech Corporation 

All rights reserved 

Compiled and edited by: 

Bob Capelli 

Stephanie Keily 

Julia Linhart 

Gerald R. Cysewski, PhD 

Editor’s Note: The publisher of this paper, Cyanotech Corporation, is the producer of 

BioAstin® Natural Astaxanthin. Cyanotech wishes to make it clear that none of the 

animal studies referenced in this paper were sponsored by Cyanotech. Our company 

policy is to sponsor medical research as human clinical trials, exclusively with subjects 

recruited as willing volunteers. We do not condone animal experimentation; yet animal 

studies done by others are reported in this paper in order that the reader may fully 

understand the ongoing medical research and the potential benefits of Astaxanthin in 

human nutrition. 

Publisher’s Note: The information contained within is for educational purposes only; it 

is not to be taken as medical advice or as an attempt to sell a particular product. People 

with medical problems or questions should consult a health professional. Information in 

this book is not intended to diagnose, treat, cure or prevent any disease. 

1

Table of Contents 

I. Introduction……………………………………………………………………3 

II. 

III. 

IV. 

Antioxidant……………………………………………………………………4 

Anti-Inflammatory: Joint, Tendon and Muscle Health….…………………..67 

Skin Health: Internal Beauty and UV Protection……………………………85 

V. Eye Health……………………………………………..……………………102 

VI. 

VII. 

Neuroprotective (Brain)…………………………………………………….128 

Cardioprotective (Heart)……………………………………………….…...149 

VIII. Immunity……………………………………………………………………198 

IX. 

Cancer Prevention and Tumor Reduction…………………………………..217 

X. Diabetes……………………………………………………………………..253 

XI. 

XII. 

Ulcers and Gastrointestinal Health……………………………………........269 

Applications for Athletes: Strength, Endurance, Energy, Recovery………281 

XIII. Additional Areas of Astaxanthin Research……………………………..…..296 

 

 

 

 

 

 

 

 

 

 

 

 

 

General Health 

Weight Loss 

Hepatoprotective (Liver) 

Renal Protective (Kidney) 

Prostate Health 

Male Fertility 

Asthma 

Candida 

Toxicity 

Neural Stem Cells 

Bioavailability 

Anti-Aging 

Safety 

2

Introduction 

The body of medical research on Astaxanthin is fast approaching critical mass for 

several diverse applications. Over the last decade in particular, the amount of studies 

done by private researchers and universities throughout the world has escalated. The 

intense interest in undertaking new research on Astaxanthin is a direct result of the 

remarkable qualities of this fascinating molecule. 

Cyanotech Corporation* feels that it is important to have a library of this research 

available for interested persons; hence we have created this document. Below the 

reader will find selected research abstracts on the health benefits of Astaxanthin. It 

was not practical to include full studies due to the substantial amount of literature 

available; however, with these abstracts, the reader will obtain a working knowledge 

of potential applications for Astaxanthin in human nutrition. The abstracts are 

presented according to health benefit as noted in the table of contents. In the case of 

studies that focused on more than one health benefit, the study is categorized 

according to the primary area of research within the abstract. 

Any questions may be directed to Cyanotech Corporation, Kailua-Kona, Hawaii, 

USA, by e-mail at info@cyanotech.com or by telephone at 808.326.1353. 

* Cyanotech Corporation is the world leader in microalgae technology. 

Cyanotech produces BioAstin Natural Astaxanthin at its 90 acre (40 hectare) 

microalgae farm on the pristine Kona Coast of Hawaii. 

3

Antioxidant 

Carotenoid Science, Vol.11, 2007, 16-20 

Quenching Activities of Common Hydrophilic and Lipophilic 

Antioxidants against Singlet Oxygen Using Chemiluminescence 

Detection System 

Yasuhiro Nishida*, Eiji Yamashita and Wataru Miki 

Institute for Food Science Research, Japan 

The singlet oxygen quenching activities among common hydrophilic and 

lipophilic antioxidants such as polyphenols, tocopherols, carotenoids, ascorbic 

acid, coenzyme Q10 and α-lipoic acid were recorded under the same test 

condition: the chemiluminescence detection system for direct 1O2 counting using 

the thermodissociable endoperoxides of 1,4-dimethylnaphthalene as 1O2 

generator in DMF : CDCl3 (9 : 1). Carotenoids exhibited larger total quenching 

rate constants than other antioxidants, with astaxanthin showing the strongest 

activity. α-Tocopherol and α-lipoic acid showed considerable activities, whereas 

the activities of ascorbic acid, CoQ10 and polyphenols were only slight; these 

included capsaicin, probucol, edaravon, BHT and Trolox. This system has the 

potential of being a powerful tool to evaluate the quenching activity against 

singlet oxygen for various hydrophilic and lipophilic compounds. 

Antioxidant 

4

Adapted from Nishida, Yamashita, Miki, Carotenoid Science, Vol. 

11, 2007, 16-20 (in Japanese) 

Astaxanthin has exceptional antioxidant activity to combat singlet oxygen when 

compared to other antioxidants. In particular, Astaxanthin can be used to defend 

against singlet oxygen damage for eye and skin health, which are especially 

susceptible to UV damage and aging effects. 

Singlet oxygen is an active oxygen species generated in human skin by exposure to 

ultraviolet radiation (UV) that causes skin damage and eye damage. In this study, 

Astaxanthin extracted from Haematococcus microalgae powerfully quenched singlet 

oxygen. Results show that the quenching effect of Astaxanthin is 800 times greater 

than coenzyme Q10. Astaxanthin was also about 75 times greater than alpha lipoic 

acid, about 550 times greater than green tea catechins and about 6000 times greater 

than Vitamin C. 

Antioxidant 

5

Carotenoids as Singlet Oxygen Quenchers in Marine 

Organisms 

Shimidzu, Gogo, Miki, 1995. Fisheries Science 62(1), 134-137 

Results indicated that Astaxanthin was significantly stronger than all other antioxidants 

tested as singlet oxygen quenchers. Among the results Astaxanthin was shown to be 

550X stronger than Vitamin E; 11X stronger than Beta-Carotene; 2.75X stronger than 

Lutein. 

Antioxidant 

6

OXYGEN FREE RADICAL SCAVENGING ABILITIES OF 

VITAMINS C, E, -CAROTENE, PYCNOGENOL, GRAPE SEED 

PROANTHOCYANIDIN EXTRACT AND ASTAXANTHINS IN 

VITRO 

Debasis Bagchi, Ph.D. Pharmacy Sciences, Creighton University School of Health 

Sciences, June 2001 

Summary: Natural Astaxanthin (as BioAstin® from Cyanotech) showed significantly 

higher free radical scavenging activity than all other antioxidants tested. Results on a 

pure active basis were as follows: 

Natural Astaxanthin Alternate Antioxidant Multiple of Greater Free 

Radical Scavenging Activity 

BioAstin Vitamin C 65X stronger 

BioAstin Vitamin E 14X stronger 

BioAstin Beta Carotene 54X stronger 

BioAstin Pycnogenol® 18X stronger 

BioAstin Synthetic Astaxanthin 21X stronger 

Antioxidant 

7

Comparison of Astaxanthin’s Singlet Oxygen Quenching Activity 

with Common Fat and Water Soluble Antioxidants 

United States Patent Application 20060217445 

Kind Code 

A1 

Chew; Boon P. ; et al. September 28, 2006 

Natural astaxanthin extract reduces DNA oxidation 

Abstract 

Provided herein are methods for reducing oxidative DNA damage in a subject, by 

administering to the subject astaxanthin, for instance a natural, astaxanthinenriched 

extract from Haematococcus pluvialis. It is shown that doses as low as 2 

mg/day, given orally to a human subject for a period of four weeks, is sufficient to 

reduced measurable endogenous oxidative DNA damage by about 40%. 

Antioxidant 

8

Phytother Res. 2009 Jun 22. [Epub ahead of print] 

Cytoprotective role of astaxanthin against glycated protein/iron 

chelate-induced toxicity in human umbilical vein endothelial cells. 

Nishigaki I, Rajendran P, Venugopal R, Ekambaram G, Sakthisekaran D, 

Nishigaki Y. 

NPO International Laboratory of Biochemistry, 1-166 Uchide, Nakagawa-ku 

Nagoya 454-0926, Japan. 

Astaxanthin (ASX), a red carotenoid pigment with no pro-vitamin A activity, is a 

biological antioxidant that occurs naturally in a wide variety of plants, algae and 

seafoods. This study investigated whether ASX could inhibit glycated protein/iron 

chelate-induced toxicity in human umbilical-vein endothelial cells (HUVEC) by 

interfering with ROS generation in these cells. Glycated fetal bovine serum 

(GFBS) was prepared by incubating fetal bovine serum (FBS) with highconcentration 

glucose. Stimulation of cultured HUVECs with 50 mm 1 mL of 

GFBS significantly enhanced lipid peroxidation and decreased antioxidant 

enzyme activities and levels of phase II enzymes. However, preincubation of the 

cultures with ASX resulted in a marked decrease in the level of lipid peroxide 

(LPO) and an increase in the levels of antioxidant enzymes in an ASX 

concentration-dependent manner. These results demonstrate that ASX could 

inhibit LPO formation and enhance the antioxidant enzyme status in GFBS/iron 

chelate-exposed endothelial cells by suppressing ROS generation, thereby 

limiting the effects of the AGE-RAGE interaction. The results indicate that ASX 

could have a beneficial role against glycated protein/iron chelate-induced toxicity 

by preventing lipid and protein oxidation and increasing the activity of 

antioxidant enzymes. 

PMID: 19548280 [PubMed - as supplied by publisher] 

Antioxidant 

9

Biochim Biophys Acta. 2001 Jun 6;1512(2):251-8. 

Efficient radical trapping at the surface and inside the phospholipid 

membrane is responsible for highly potent antiperoxidative activity 

of the carotenoid astaxanthin. 

Goto S, Kogure K, Abe K, Kimata Y, Kitahama K, Yamashita E, Terada H. 

Faculty of Pharmaceutical Sciences, University of Tokushima, Japan. 

Kogure@ph.tokushima-u.ac.jp 

The effects of the carotenoids beta-carotene and astaxanthin on the peroxidation 

of liposomes induced by ADP and Fe(2+) were examined. Both compounds 

inhibited production of lipid peroxides, astaxanthin being about 2-fold more 

effective than beta-carotene. The difference in the modes of destruction of the 

conjugated polyene chain between beta-carotene and astaxanthin suggested that 

the conjugated polyene moiety and terminal ring moieties of the more potent 

astaxanthin trapped radicals in the membrane and both at the membrane surface 

and in the membrane, respectively, whereas only the conjugated polyene chain of 

beta-carotene was responsible for radical trapping near the membrane surface and 

in the interior of the membrane. The efficient antioxidant activity of astaxanthin is 

suggested to be due to the unique structure of the terminal ring moiety. 

Publication Types: 

PMID: 11406102 [PubMed - indexed for MEDLINE] 

Antioxidant 

10

Chem Biol Interact. 2009 Aug 14;180(3):398-406. Epub 2009 Apr 2. 

Intervention of astaxanthin against cyclophosphamide-induced 

oxidative stress and DNA damage: a study in mice. 

Tripathi DN, Jena GB. 

Department of Pharmacology and Toxicology, National Institute of 

Pharmaceutical Education and Research, Sector-67, S.A.S. Nagar, Punjab 

160062, India. 

Astaxanthin, a natural and nutritional red carotenoid pigment, is used as a dietary 

supplement. The intention of the present study was to investigate the beneficial 

effects of dietary pigment astaxanthin, against cyclophosphamide-induced 

oxidative stress and DNA damage. The end points of evaluation of the study 

included: (a) malondialdehyde, glutathione and superoxide dismutase 

concentration in liver to detect oxidative stress; (b) normal and modified alkaline 

comet assays (the latter includes lesion-specific enzymes formamidopyrimidine- 

DNA glycosylase and endonuclease-III) to detect normal and oxidative stressinduced 

DNA damage by cyclophosphamide in the mouse bone marrow and the 

peripheral blood lymphocytes. In addition, micronucleus assay and chromosomal 

aberration test capable of detecting the DNA damage were also carried out in 

peripheral blood and bone marrow of mice. Cyclophosphamide (100 mg/kg intraperitoneal) 

treatment led to significant increase in liver malondialdehyde and 

decreased the antioxidant enzymes glutathione and superoxide dismutase. Further, 

cyclophosphamide also significantly increased the DNA damage as observed 

from normal and modified comet assays as well as micronucleus and 

chromosomal aberration assay. Pre-treatment with astaxanthin (12.5, 25 and 50 

mg/kg/day for 5 days per oral) resulted in the restoration of oxidative stress 

markers such as malondialdehyde, glutathione and superoxide dismutase in liver. 

The amelioration of oxidative stress with astaxanthin pre-treatment correlated 

well with the decreased DNA damage as evident from normal and modified 

alkaline comet assays of bone marrow cells and peripheral blood lymphocytes. 

Further astaxanthin pre-treatment also reduced the frequency of chromosomal 

breakage and micronucleus formation in the mouse bone marrow cells and 

peripheral blood reticulocytes. It is thus concluded that pre-treatment with 

astaxanthin attenuates cyclophosphamide-induced oxidative stress and subsequent 

DNA damage in mice and it can be used as a chemoprotective agent against the 

toxicity of anticancer drug cyclophosphamide. 

Research Support, Non-U.S. Gov't 

PMID: 19539803 [PubMed - in process] 

Antioxidant 

11

J Agric Food Chem. 2000 Apr;48(4):1150-4. 

Antioxidant activities of astaxanthin and related carotenoids. 

Naguib YM. 

Phytochem Technologies, Chelmsford, MA 01824, USA. 

The antioxidant activities of astaxanthin and related carotenoids have been 

measured by employing a newly developed fluorometric assay. This assay is 

based on 4,4-difluoro-3,5-bis(4-phenyl-1, 3-butadienyl)-4-bora-3a,4a-diaza-sindacene 

(BODIPY 665/676) as an indicator; 2,2'-azobis-2,4- 

dimethylvaleronitrile (AMVN) as a peroxyl radical generator; and 6-hydroxy- 

2,5,7, 8-tetramethylchroman-2-carboxylic acid (Trolox) as a calibrator in an 

organic and liposomal media. By employing this assay, three categories of 

carotenoids were examined: namely, the hydrocarbon carotenoids lycopene, 

alpha-carotene, and beta-carotene; the hydroxy carotenoid lutein; and the alphahydroxy-ketocarotenoid 

astaxanthin. The relative peroxyl radical scavenging 

activities of Trolox, astaxanthin, alpha-tocopherol, lycopene, beta-carotene, 

lutein, and alpha-carotene in octane/butyronitrile (9:1, v/v) were determined to be 

1.0, 1.0, 1.3, 0.5, 0.4, 0.3, and 0.2, respectively. In dioleoylphosphatidyl choline 

(DOPC) liposomal suspension in Tri-HCl buffer (pH 7.4 at 40 degrees C), the 

relative reactivities of astaxanthin, beta-carotene, alpha-tocopherol, and lutein 

were found to be 1.00, 0.9, 0.6, and 0.6, respectively. When BODIPY 665/676 

was replaced by 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a, 4a-diaza-sindacene-3-undecanoic 

acid (BODIPY 581/591 C(11)) as an indicator, 

astaxanthin showed the highest antioxidant activity toward peroxyl radicals. The 

relative reactivities of Trolox, astaxanthin, alpha-tocopherol, alpha-carotene, 

lutein, beta-carotene, and lycopene were determined to be 1.0, 1.3, 0.9, 0.5, 0.4, 

0.2, and 0.4, respectively. 


Antioxidant 

12

J Nutr Biochem. 2009 May 6. [Epub ahead of print] 

Astaxanthin protects mitochondrial redox state and functional 

integrity against oxidative stress. 

Wolf AM, Asoh S, Hiranuma H, Ohsawa I, Iio K, Satou A, Ishikura M, Ohta 

S. 

Department of Biochemistry and Cell Biology, Institute of Development and 

Aging Sciences, Nippon Medical School, Nakahara-ku, Kawasaki, Kanagawa 

211-8533, Japan. 

Mitochondria combine the production of energy with an efficient chain of 

reduction-oxidation (redox) reactions but also with the unavoidable production of 

reactive oxygen species. Oxidative stress leading to mitochondrial dysfunction is 

a critical factor in many diseases, such as cancer and neurodegenerative and 

lifestyle-related diseases. Effective antioxidants thus offer great therapeutic and 

preventive promise. Investigating the efficacy of antioxidants, we found that a 

carotenoid, astaxanthin (AX), decreased physiologically occurring oxidative 

stress and protected cultured cells against strong oxidative stress induced with a 

respiratory inhibitor. Moreover, AX improved maintenance of a high 

mitochondrial membrane potential and stimulated respiration. Investigating how 

AX stimulates and interacts with mitochondria, a redox-sensitive fluorescent 

protein (roGFP1) was stably expressed in the cytosol and mitochondrial matrix to 

measure the redox state in the respective compartments. AX at nanomolar 

concentrations was effective in maintaining mitochondria in a reduced state. 

Additionally, AX improved the ability of mitochondria to remain in a reduced 

state under oxidative challenge. Taken together, these results suggest that AX is 

effective in improving mitochondrial function through retaining mitochondria in 

the reduced state. 


Antioxidant 

13

Zhongguo Gu Shang. 2008 Mar;21(3):187-9. 

[Effects of Astaxanthin on the damage of osteoblast induced by H2O2] 

[Article in Chinese] 

Pei LP, Dong FH, Hui BD. 

Institute of Orthopaedics and Traumatology, China Academy of Chinese Medical 

Science, Beijing 100700, China. 

OBJECTIVE: To investigate the effect of Astaxanthin on enhancing the function of antioxidative 

damage in osteoblast. METHODS: MC3T3-E1 osteoblasts were randomly 

divided into five groups, including control group, model group, Astaxanthin group [lowdose 

(1 x 10(-7) mol/L), middle-dose (1 x 10(-6) mol/L), high-dose (1 x 10(-5) mol/L)], 

in which the activity of cells, activity of superoxide dismutase (SOD), the content of 

reactive oxygen species (ROS), lipid oxygen (LPO) and membrane fluidity were tested 

and compared. RESULTS: Compared with Astaxanthin groups, the activity of cells, SOD 

activity and membrane fluidity in the model group were significantly decreased (P < 

0.01). However, the contents of ROS and LPO were significantly raised (P < 0.01). 

CONCLUSION: H2O2 can cause oxidative damage of MC3T3-E1 osteoblasts, but 

Astaxanthin can prevent or decrease its influence. 


Antioxidant 

14

Phys Chem A. 2008 Sep 25;112(38):9037-42. Epub 2008 Aug 21. 

Donator acceptor map for carotenoids, melatonin and vitamins. 

Martínez A, Rodríguez-Gironés MA, Barbosa A, Costas M. 

Instituto de Investigaciones en Materiales, Universidad Nacional Autonoma de 

Mexico, Circuito Interior, S N, Ciudad Universitaria, P. O. Box 70-360, 

Coyoacan, 04510, Mexico. martina@iim.unam.mx 

Bright yellow and red colors in animals and plants are assumed to be caused by 

carotenoids (CAR). In animals, these pigments are deposited in scales, skin and 

feathers. Together with other naturally occurring and colorless substances such as 

melatonin and vitamins, they are considered antioxidants due to their free-radicalscavenging 

properties. However, it would be better to refer to them as 

"antiradicals", an action that can take place either donating or accepting electrons. 

In this work we present quantum chemical calculations for several CAR and some 

colorless antioxidants, such as melatonin and vitamins A, C and E. The antiradical 

capacity of these substances is determined using vertical ionization energy (I), 

electron affinity (A), the electrodonating power (omega(-)) and the 

electroaccepting power (omega(+)). Using fluor and sodium as references, 

electron acceptance (R(a)) and electron donation (R(d)) indexes are defined. A 

plot of R(d) vs R(a) provides a donator acceptor map (DAM) useful to classify 

any substance regarding its electron donating-accepting capability. Using this 

DAM, a qualitative comparison among all the studied compounds is presented. 

According to R(d) values, vitamin E is the most effective antiradical in terms of 

its electron donor capacity, while the most effective antiradical in terms of its 

electron acceptor capacity, R(a), is astaxanthin, the reddest CAR. These results 

may be helpful for understanding the role played by naturally occurring pigments, 

acting as radical scavengers either donating or accepting electrons. 


 



Antioxidant 

15

Food Chem Toxicol. 2008 Jan;46(1):212-9. Epub 2007 Aug 14. 

Effect of astaxanthin on kidney function impairment and oxidative 

stress induced by mercuric chloride in rats. 

Augusti PR, Conterato GM, Somacal S, Sobieski R, Spohr PR, Torres JV, 

Charão MF, Moro AM, Rocha MP, Garcia SC, Emanuelli T. 

Post-graduate Program on Toxicological Biochemistry, Center of Natural and 

Exact Sciences, Federal University of Santa Maria, 97105-900 Santa Maria, RS, 

Brazil. 

Reactive oxygen species are implicated as mediators of tissue damage in the acute 

renal failure induced by inorganic mercury. Astaxanthin (ASX), a carotenoid with 

potent antioxidant properties, exists naturally in various plants, algae, and 

seafoods. This paper evaluated the ability of ASX to prevent HgCl(2) 

nephrotoxicity. Rats were injected with HgCl(2) (0 or 5 mg/kg b.w., sc) 6h after 

ASX had been administered (0, 10, 25, or 50mg/kg, by gavage) and were killed 

12h after HgCl(2) exposure. Although ASX prevented the increase of lipid and 

protein oxidation and attenuated histopathological changes caused by HgCl(2) in 

kidney, it did not prevent creatinine increase in plasma and delta-aminolevulinic 

acid dehydratase inhibition induced by HgCl(2). Glutathione peroxidase and 

catalase activities were enhanced, while superoxide dismutase activity was 

depressed in HgCl(2)-treated rats when compared to control and these effects 

were prevented by ASX. Our results indicate that ASX could have a beneficial 

role against HgCl(2) toxicity by preventing lipid and protein oxidation, changes in 

the activity of antioxidant enzymes and histopathological changes. 



Antioxidant 

16

Biochem Biophys Res Commun. 2007 May 25;357(1):187-93. Epub 2007 Mar 28. 

Cis astaxanthin and especially 9-cis astaxanthin exhibits a higher 

antioxidant activity in vitro compared to the all-trans isomer. 

Liu X, Osawa T. 

Laboratory of Food and Biodynamics, Graduate School of Bioagricultural 

Science, Nagoya University, Nagoya 464-8601, Japan. 

In recent years, a number of studies have implicated the potent antioxidant 

property of astaxanthin in various experimental systems; however, these studies 

employed only the all-trans isomer. On the other hand, it has been reported that 

all-trans natural astaxanthin is readily isomerized to cis-trans, especially 9-cis and 

13-cis isomers, under certain conditions by chemical analysis; however, the 

biological activities of the cis isomers of astaxanthin are little known. In the 

present study, we investigated the antioxidant activity of 9-cis and 13-cis 

astaxanthin compared to the all-trans isomer in vitro. In a stable radical DPPH 

scavenging activity test and in rat microsome and rabbit erythrocyte ghost 

membrane lipid peroxidation systems induced by AAPH and t-BuOOH, 

respectively, the results apparently showed that cis-astaxanthin, especially 9-cis 

astaxanthin, exhibited a higher antioxidant effect than the all-trans isomer. In 

addition, during polyunsaturated fatty acid (PUFA) oxidation, both DHA and 

linoleic acid hydroperoxides formation were markedly inhibited by astaxanthin 

isomers addition in the order 9-cis >13-cis >all-trans. Furthermore, 9-cis also 

exhibited the most effective inhibition of the generation of ROS induced by 6- 

hydroxydopamine (6-OHDA) in human neuroblastoma SH-SY5Y cells among the 

astaxanthin isomers, as well as on the degradation of collagen type II induced by 

DHA and linoleic acid hydroperoxides. The above-mentioned results suggest, for 

the first time, that cis isomer astaxanthin, especially 9-cis astaxanthin, has a much 

higher antioxidant potency than that of the all-trans isomer. 


Antioxidant 

17

Biochim Biophys Acta. 2007 Jan;1768(1):167-74. Epub 2006 Sep 22. 

Differential effects of carotenoids on lipid peroxidation due to 

membrane interactions: X-ray diffraction analysis. 

McNulty HP, Byun J, Lockwood SF, Jacob RF, Mason RP. 

Elucida Research, 100 Cummings Center, Suite 135L, P.O. Box 7100, Beverly, 

MA 01915-0091, USA. hmcnulty@elucidaresearch.com 

The biological benefits of certain carotenoids may be due to their potent 

antioxidant properties attributed to specific physico-chemical interactions with 

membranes. To test this hypothesis, we measured the effects of various 

carotenoids on rates of lipid peroxidation and correlated these findings with their 

membrane interactions, as determined by small angle X-ray diffraction 

approaches. The effects of the homochiral carotenoids (astaxanthin, zeaxanthin, 

lutein, beta-carotene, lycopene) on lipid hydroperoxide (LOOH) generation were 

evaluated in membranes enriched with polyunsaturated fatty acids. Apolar 

carotenoids, such as lycopene and beta-carotene, disordered the membrane bilayer 

and showed a potent pro-oxidant effect (>85% increase in LOOH levels) while 

astaxanthin preserved membrane structure and exhibited significant antioxidant 

activity (40% decrease in LOOH levels). These findings indicate distinct effects 

of carotenoids on lipid peroxidation due to membrane structure changes. These 

contrasting effects of carotenoids on lipid peroxidation may explain differences in 

their biological activity. 


Antioxidant 

18

Luminescence. 2005 Nov-Dec;20(6):419-27. 

Comparative study of antioxidants as quenchers or scavengers of 

reactive oxygen species based on quenching of MCLA-dependent 

chemiluminescence. 

Hosaka S, Obuki M, Nakajima J, Suzuki M. 

Department of Applied Chemistry, Faculty of Engineering, Tokyo Polytechnic 

University, 1583 Atsugi, Kanagawa 243-0297, Japan. s-hosaka@parkcity.ne.jp 

The quenching or scavenging effect of non-enzymatic antioxidants against 

reactive oxygen species (ROS) was studied by comparing the degree of 

suppression of chemiluminescence (CL) caused by the oxidation of MCLA 

(methoxylated Cypridina luciferin analogue) by ROS. MCLA-dependent CL 

caused by O2- was effectively quenched by ascorbic acid, beta-carotene, lycopene 

and astaxanthin, while it was enhanced by alpha-tocopherol. The CL by 1O2 was 

quenched effectively by beta-carotene, lycopene and astaxanthin, moderately by 

ascorbic acid, and slightly by alpha-tocopherol. beta-Carotene and alphatocopherol 

remarkably suppressed the CL when ROS was HO*. The present study 

revealed that MCLA-dependent CL assay provides a simple and rapid method for 

the evaluation of antioxidants as a quencher or scavenger against any kind of 

ROS. (c) 2005 John Wiley & Sons, Ltd. 



Antioxidant 

19

Bioorg Med Chem Lett. 2004 Aug 2;14(15):3985-91. 

Synthesis, characterization, and direct aqueous superoxide anion 

scavenging of a highly water-dispersible astaxanthin-amino acid 

conjugate. 

Jackson HL, Cardounel AJ, Zweier JL, Lockwood SF. 

Hawaii Biotech, Inc., 99-193 Aiea Heights Drive, Suite 200, Aiea, HI 96701, 

USA. 

The aqueous solubility and/or dispersibility of synthetic carotenoid analogs can be 

improved by varying the chemical structure(s) of the esterified moieties. In the 

current study, a highly water-dispersible astaxanthin (3,3'-dihydroxy-beta,betacarotene-4,4'-dione) 

derivative was synthesized by esterification to the amino acid 

L-lysine, and subsequently converted to the tetrahydrochloride salt. Deep violet, 

evenly colored aqueous suspensions were obtained with addition of the novel 

derivative to USP purified water up to a maximum of 181.6 mg/mL. These 

aqueous suspensions were obtained without the addition of heat, detergents, cosolvents, 

or other additives. At higher concentrations (above 181.6 mg/mL), the 

dispersion became turbid and viscous. There was no saturation point up to 181.6 

mg/mL. The direct superoxide scavenging ability of the tetrahydrochloride 

dilysine astaxanthin salt was also evaluated by electron paramagnetic resonance 

(EPR) spectroscopy in a well-characterized in vitro isolated human neutrophil 

assay. The novel derivative was an extremely potent (micromolar concentration) 

aqueous-phase scavenger, with near-complete suppression of the superoxide 

anion signal (as detected by spin-trap adducts of DEPMPO) achieved at 100 

microM. To the authors' knowledge, this novel carotenoid derivative exhibits the 

greatest aqueous dispersibility yet described for a natural and/or synthetic C40 

carotenoid, and as such, will find utility in those applications for which aqueousphase 

singlet oxygen quenching and direct radical scavenging are required. 


Antioxidant 

20

Chem Phys Lipids. 2001 Nov;113(1-2):11-22. 

Molecular characteristics of astaxanthin and beta-carotene in the 

phospholipid monolayer and their distributions in the phospholipid 

bilayer. 

Shibata A, Kiba Y, Akati N, Fukuzawa K, Terada H. 

Faculty of Pharmaceutical Sciences, The University of Tokushima, Shomachi, 

Tokushima 770-8505, Japan. ashibata@ph.tokushima-u.ac.jp 

The molecular characteristics of the monolayers of astaxanthin with polar group 

on the beta-ionone ring in the molecule and beta-carotene without polar group and 

their interactions in mixed carotenoid-phospholipid monolayers and the effects of 

carotenoids on the phase behavior of the phospholipid bilayers were examined by 

the monolayer technique and differential scanning calorimetry (DSC). We found 

from the monolayer study that beta-carotene had an amphiphilic nature. The 

molecular assembly of astaxanthin in the monolayer at the 

hydrophobic/hydrophilic interface was more stable than that of beta-carotene. 

Dimyristoylphosphatidylcholine (DMPC) in the monolayer was miscible with 

astaxanthin in the range of 0-0.4 mol fractions of astaxanthin, but not fully 

miscible with beta-carotene even at low concentrations below 0.1 mol fraction of 

beta-carotene. Surface potential and compression/expansion cycles of betacarotene 

monolayer indicated the formation of molecular aggregates by itself. 

DSC study showed that when small amount of astaxanthin was added, the 

transition temperature of dipalmitoylphosphatidylcholine (DPPC) was markedly 

shifted to lower temperatures and that the transition peak was asymmetrically 

broadened, indicative of a significant depression in cooperativity of the gel to 

liquid-crystalline transition. The asymmetric DSC endothermic bands of DPPC 

incorporating small amounts of astaxanthin were well fit by deconvolution into 

two to three domains containing different concentrations of astaxanthin. On the 

contrary, the incorporation of beta-carotene resulted in a small depression of the 

main transition temperature with a slight broadening of the transition peak, 

suggesting a small miscibility of beta-carotene with the phospholipid bilayer or a 

formation of aggregates of beta-carotene in the membranes. These results suggest 

that there would be a high localized concentration in the phase separated 

membrane for astaxanthin or beta-carotene to function effectively as scavenger. 


Antioxidant 

21

Biochem Biophys Res Commun. 2001 Oct 19;288(1):225-32. 

Astaxanthin and peridinin inhibit oxidative damage in Fe(2+)- 

loaded liposomes: scavenging oxyradicals or changing membrane 

permeability? 

Barros MP, Pinto E, Colepicolo P, Pedersén M. 

Department of Botany, Stockholm University, SE-10691 Stockholm, Sweden. 

mpbarros@botan.su.se 

Astaxanthin and peridinin, two typical carotenoids of marine microalgae, and 

lycopene were incorporated in phosphatidylcholine multilamellar liposomes and 

tested as inhibitors of lipid oxidation. Contrarily to peridinin results, astaxanthin 

strongly reduced lipid damage when the lipoperoxidation promoters-H(2)O(2), 

tert-butyl hydroperoxide (t-ButOOH) or ascorbate-and Fe(2+):EDTA were added 

simultaneously to the liposomes. In order to check if the antioxidant activity of 

carotenoids was also related to their effect on membrane permeability, the 

peroxidation processes were initiated by adding the promoters to Fe(2+)-loaded 

liposomes (encapsulated in the inner aqueous solution). Despite that the 

rigidifying effect of carotenoids in membranes was not directly measured here, 

peridinin probably has decreased membrane permeability to initiators (t-ButOOH 

> ascorbate > H(2)O(2)) since its incorporation limited oxidative damage on ironliposomes. 

On the other hand, the antioxidant activity of astaxanthin in ironcontaining 

vesicles might be derived from its known rigidifying effect and the 

inherent scavenging ability. 



Antioxidant 

22

Arch Biochem Biophys. 2001 Jan 1;385(1):13-9. 

The interaction of dietary carotenoids with radical species. 

Mortensen A, Skibsted LH, Truscott TG. 

Department of Dairy and Food Science, Royal Veterinary and Agricultural 

University, Frederiksberg, Denmark. 

Dietary carotenoids react with a wide range of radicals such as CCl3O2*, RSO2*, 

NO2*, and various arylperoxyl radicals via electron transfer producing the radical 

cation of the carotenoid. Less strongly oxidizing radicals, such as alkylperoxyl 

radicals, can lead to hydrogen atom transfer generating the neutral carotene 

radical. Other processes can also arise such as adduct formation with sulphurcentered 

radicals. The oxidation potentials have been established, showing that, in 

Triton X-100 micelles, lycopene is the easiest carotenoid to oxidize to its radical 

cation and astaxanthin is the most difficult. The interaction of carotenoids and 

carotenoid radicals with other antioxidants is of importance with respect to antiand 

possibly pro-oxidative reactions of carotenoids. In polar environments the 

vitamin E (alpha-tocopherol) radical cation is deprotonated (TOH*+ --> TO* + 

H+) and TO* does not react with carotenoids, whereas in nonpolar environments 

such as hexane, TOH*+ is converted to TOH by hydrocarbon carotenoids. 

However, the nature of the reaction between the tocopherol and various 

carotenoids shows a marked variation depending on the specific tocopherol 

homologue. The radical cations of the carotenoids all react with vitamin C so as to 

"repair" the carotenoid. 


 

 


Review 


Antioxidant 

23

J Nutr. 2000 Jul;130(7):1800-8. 

Depletion of alpha-tocopherol and astaxanthin in Atlantic salmon 

(Salmo salar) affects autoxidative defense and fatty acid metabolism. 

Bell JG, McEvoy J, Tocher DR, Sargent JR. 

Institute of Aquaculture, University of Stirling, Stirling FK9 4LA, Scotland, U.K. 

Duplicate groups of Atlantic salmon post-smolts were fed four purified diets 

supplemented with both vitamin E and the carotenoid astaxanthin (Ax) (+E, +Ax), 

or supplemented with either vitamin E or Ax (-E, +Ax and +E, -Ax) or deficient 

in both vitamin E and Ax (-E, -Ax) for 22 wk. There were no effects of diet on 

growth rate, but an extensive lipoid liver degenerative lesion was observed in 

15% of fish fed diets deficient in vitamin E. Tissue vitamin E concentrations 

varied in accordance with dietary vitamin E in liver, muscle, heart, plasma, brain 

and eye; levels were reduced to approximately 3% in liver but only to 40% in eye 

of fish fed diets deficient in vitamin E compared with those fed diets 

supplemented with vitamin E. An interactive sparing of Ax supplementation on 

tissue vitamin E concentration was observed, but only in brain. Dietary deficiency 

of both vitamin E and Ax significantly increased the recovery of desaturated and 

elongated products of both [1-(14)C] 18:3(n-3) and [1-(14)C] 20:5(n-3) in isolated 

hepatocytes, suggesting that conversion of fatty acids to their long-chain highly 

unsaturated products can be stimulated by a deficiency of lipid-soluble 

antioxidants. The antioxidant synergism of vitamin E and Ax was supported by 

their ability to reduce malondialdehyde formation in an in vitro stimulation of 

microsomal lipid peroxidation and to reduce plasma levels of 8-isoprostane. The 

results of this study suggest that both vitamin E and the carotenoid Ax have 

antioxidant functions in Atlantic salmon. 



Antioxidant 

24

Biochim Biophys Acta. 2000 Jan 15;1463(1):179-87. 

Exogenously incorporated ketocarotenoids in large unilamellar 

vesicles. Protective activity against peroxidation. 

Rengel D, Díez-Navajas A, Serna-Rico A, Veiga P, Muga A, Milicua JC. 

Department of Biochemistry and Molecular Biology, University of the Basque 

Country, P.O. Box 644, 48080, Bilbao, Spain. 

The ability of astaxanthin and canthaxanthin as chain-breaking antioxidants was 

studied in Cu(2+)-initiated peroxidation of phosphatidylcholine large unilamellar 

vesicles (LUVs). Both carotenoids increased the lag period that precedes the 

maximum rate of lipid peroxidation, though astaxanthin showed stronger activity. 

For these experiments, different amounts of xanthophylls were exogenously 

added to previously made LUVs, non-incorporated pigment being afterwards 

removed. Differential scanning calorimetry assays with L-beta,gammadimyristoyl-alpha-phosphatidylcholine 

LUVs demonstrated that xanthophylls 

incorporated as described interact with the lipid matrix becoming interspersed 

among the phospholipid molecules. 



Antioxidant 

25

FEBS Lett. 1997 Nov 24;418(1-2):91-7. 

Comparative mechanisms and rates of free radical scavenging by 

carotenoid antioxidants. 

Mortensen A, Skibsted LH, Sampson J, Rice-Evans C, Everett SA. 

Department of Dairy and Food Science, Royal Veterinary and Agricultural 

University, Frederiksberg C, Denmark. 

The comparative mechanisms and relative rates of nitrogen dioxide (NO2.), thiyl 

(RS.) and sulphonyl (RSO2.) radical scavenging by the carotenoid antioxidants 

lycopene, lutein, zeaxanthin, astaxanthin and canthaxanthin have been determined 

by pulse radiolysis. All the carotenoids under study react with the NO2. radical 

via electron transfer to generate the carotenoid radical cation (Car.+). In marked 

contrast the glutathione and 2-mercaptoethanol thiyl radicals react via a radical 

addition process to generate carotenoid-thiyl radical adducts [RS-Car].. The 

RSO2. radical undergoes both radical addition, [RSO2-Car]. and electron 

abstraction, Car.+. Both carotenoid adduct radicals and radical cations decay 

bimolecularly. Absolute rate constants for radical scavenging were in the order of 

approximately 10(7)-10(9) M(-1) s(-1) and follow the sequence HO(CH2)2S. > 

RSO2. > GS. > NO2.. Although there were some discernible trends in carotenoid 

reactivity for individual radicals, rate constants varied by no greater than a factor 

of 2.5. The mechanism and rate of scavenging is strongly dependent on the nature 

of the oxidising radical species but much less dependent on the carotenoid 

structure. 



Antioxidant 

26

J Nutr Sci Vitaminol (Tokyo). 1997 Jun;43(3):345-55. 

Inhibition of beta-carotene and astaxanthin of NADPH-dependent 

microsomal phospholipid peroxidation. 

Nakagawa K, Kang SD, Park DK, Handelman GJ, Miyazawa T. 

Department of Applied Biological Chemistry, Tohoku University, Sendai, Japan. 

To evaluate the antioxidant effects of beta-carotene and astaxanthin, rat liver 

microsomes were exposed to a mixture of chelated iron (Fe3+/ADP) and 

NADPH. The carotenoids (190 pmol/mg protein) were incorporated into some of 

these microsomal membranes, and phospholipid hydroperoxides (PLOOH), 

thiobarbituric acid reactive substances (TBARS) and endogenous alphatocopherol 

content were measured over time after the initiation of oxidant stress. 

In control microsomes, oxidant stress led to accumulation of 1,865 (+/- 371) pmol 

PLOOH/mg protein during the initial 10-min peroxidation reaction, followed by a 

more gradual decrease during the subsequent 20-min of reaction. PLOOH 

accumulation during the initial 10-min reaction period was reduced to 588 (+/- 

169) pmol/mg protein with beta-carotene present and 800 (+/- 288) pmol/mg 

protein with astaxanthin present. During the following 20-min of incubation, 

PLOOH levels declined in the carotenoid-supplemented microsomes but 

continued to increase at a slower rate in control preparations. TBARS did not 

show such large accumulation as observed in PLOOH during the initial 10-min 

incubation in any microsomal sample. The presence of carotenoids in the 

microsomal membrane partially inhibited the loss of alpha-tocopherol, especially 

during the later phase of oxidant stress. When lipid peroxidation is generated by 

membrane-bound cyt-P450, the specific measurement of PLOOH clearly 

demonstrates that the presence of carotenoids provides antioxidant protection. 


Antioxidant 

27

Z Lebensm Unters Forsch. 1993 May;196(5):423-9. 

Carotenoid scavenging of radicals. Effect of carotenoid structure 

and oxygen partial pressure on antioxidative activity. 

Jørgensen K, Skibsted LH. 

RVAU Centre for Food Research, Royal Veterinary and Agricultural University, 

Frederiksberg, Denmark. 

Carotenoid scavenging of free radicals has been investigated in peroxidizing 

methyl esters of unsaturated fatty acids using (i) metmyoglobin as a water-based 

free-radical initiator in a heterogeneous lipid/water system, and (ii) azo-bisisobutyronitrile 

as a free-radical initiator in a homogeneous chloroform solution. 

For the heterogeneous system, using a combination of electrochemical oxygen 

depletion measurements, spectrophotometric determination of lipid 

hydroperoxides and carotenoid degradation, it was demonstrated that each of the 

four carotenoids astaxanthin, beta-carotene, canthaxanthin, and zeaxanthin 

protects the methyl esters against oxidation. The antioxidative effect increases 

with increasing carotenoid concentration, increases with decreasing oxygen 

partial pressure (0.010 < pO2 < 0.50 atm), and shows little dependence on the 

structure of the carotenoid. For a homogeneous solution, the effect of the structure 

of the carotenoid was further investigated, and it was shown that the stability of 

the four carotenoids in the oxidizing system are different, with the order of 

decreasing stability being: astaxanthin > canthaxanthin > beta-carotene > 

zeaxanthin. Each of the four carotenoids can suppress lipid oxidation and the 

degree of suppression of peroxidation of methyl linoleate corresponds to the 

difference in stability. 


Antioxidant 

28

Arch Biochem Biophys. 1992 Sep;297(2):291-5. 

Astaxanthin and canthaxanthin are potent antioxidants in a 

membrane model. 

Palozza P, Krinsky NI. 

Department of Biochemistry, Tufts University School of Medicine, Boston, 

Massachusetts 02111-1837. 

When the conjugated keto-carotenoids, either astaxanthin or canthaxanthin, are 

added to rat liver microsomes undergoing radical-initiated lipid peroxidation 

under air, they are as effective as alpha-tocopherol in inhibiting this process. This 

contrasts with the effect of beta-carotene, which is a much less potent antioxidant 

when added in this system, without the addition of other antioxidants. 



Antioxidant 

29

Biochim Biophys Acta. 1992 Jun 22;1126(2):178-84. 

Antioxidant activity of xanthophylls on peroxyl radical-mediated 

phospholipid peroxidation. 

Lim BP, Nagao A, Terao J, Tanaka K, Suzuki T, Takama K. 

National Food Research Institute, Ministry of Agriculture, Forestry and Fisheries, 

Ibaraki, Japan. 

The ability of xanthophylls (canthaxanthin, zeaxanthin, and astaxanthin) as chainbreaking 

antioxidants was investigated in peroxyl radical-mediated peroxidation 

of phosphatidylcholine (PC) liposomes under atmospheric conditions using lipidsoluble 

and water-soluble radical generators. These xanthophylls retarded the 

chain propagation reaction of phosphatidylcholine hydroperoxides (PC-OOH) 

formation, although their activities to trap chain-carrying peroxyl radical were 

much less than that of alpha-tocopherol. In chick plasma studies, it was observed 

that endogenious xanthophylls participated in the antioxidant defenses against the 

attack of aqueous peroxyl radical. It was concluded that xanthophylls possess the 

ability to act as chain-breaking antioxidants in the peroxidation of membraneous 

phospholipids. Dietary xanthophylls may, therefore, be helpful in resisting 

membraneous phospholipids against oxidative damage in vivo. 


Antioxidant 

30

Physiol Chem Phys Med NMR. 1990;22(1):27-38. 

Inhibition of oxidative injury of biological membranes by 

astaxanthin. 

Kurashige M, Okimasu E, Inoue M, Utsumi K. 

Department of Medical Biology, Kochi Medical School, Japan. 

The value of astaxanthin, a carotenoid pigment, in the treatment of oxidative 

injury is assessed. Astaxanthin protects the mitochondria of vitamin E-deficient 

rats from damage by Fe2(+)-catalyzed lipid peroxidation both in vivo and in vitro. 

The inhibitory effect of astaxanthin on mitochondrial lipid peroxidation is 

stronger than that of alpha-tocopherol. Thin layer chromatographic analysis shows 

that the change in phospholipid components of erythrocytes from vitamin E- 

deficient rats induced by Fe2+ and Fe3(+)-xanthine/xanthine oxidase system was 

significantly suppressed by astaxanthin. Carrageenan-induced inflammation of the 

paw is also significantly inhibited by administration of astaxanthin. These data 

indicate that astaxanthin functions as a potent antioxidant both in vivo and in 

vitro. 


Antioxidant 

31

Lipids. 1989 Jul;24(7):659-61. 

Antioxidant activity of beta-carotene-related carotenoids in solution. 

Terao J. 

Research Institute for Food Science, Kyoto University, Uji, Kyoto 611, Japan. 

The effect of the antioxidant activity of beta-carotene and related carotenoids on 

the free radical-oxidation of methyl linoleate in solution was examined by 

measuring the production of methyl linoleate hydroperoxides. Canthaxanthin and 

astaxanthin which possess oxo groups at the 4 and 4'-positions in the beta-ionone 

ring retarded the hydroperoxide formation more efficiently than beta-carotene and 

zeaxanthin which possess no oxo groups. The rates of autocatalytic oxidation of 

canthaxanthin and astaxanthin were also slower than those of beta-carotene and 

zeaxanthin. These results suggest that canthaxanthin and astaxanthin are more 

effective antioxidants than beta-carotene by stabilizing the trapped radicals. 



Antioxidant 

32

Critical Reviews in Food Science and Nutrition, 46:185–196 (2006) 

Astaxanthin: A Review of its Chemistry and Applications 

I. HIGUERA-CIAPARA, L. F ÉLIX-VALENZUELA, and F. M. 

GOYCOOLEA 

Centro de Investigación en Alimentación y Desarrollo, A.C., P.O. Box 1735. 

Hermosillo, Sonora. México. 83000 

Astaxanthin is a carotenoid widely used in salmonid and crustacean aquaculture 

to provide the pink color characteristic of that species. This application has been 

well documented for over two decades and is currently the major market driver 

for the pigment. Additionally, astaxanthin also plays a key role as an intermediary 

in reproductive processes. Synthetic astaxanthin dominates the world market but 

recent interest in natural sources of the pigment has increased substantially. 

Common sources of natural astaxanthin are the green algae Haematococcus 

pluvialis, the red yeast, Phaffia rhodozyma, as well as crustacean byproducts. 

Astaxanthin possesses an unusual antioxidant activity which has caused a surge in 

the nutraceutical market for the encapsulated product. Also, health benefits such 

as cardiovascular disease prevention, immune system boosting, bioactivity against 

Helycobacter pylori, and cataract prevention, have been associated with 

astaxanthin consumption. Research on the health benefits of astaxanthin is very 

recent and has mostly been performed in vitro or at the 

pre-clinical level with humans. This paper reviews the current available evidence 

regarding astaxanthin chemistry and its potential beneficial effects in humans. 

Antioxidant 

33

Lebensm Unters Forsch (1993) 196: 423-429 

Carotenoid Scavenging of Radicals 

Effect of carotenoid structure and oxygen partial pressure on 

antioxidative activiy 

Kevin Jorgensen and Leif H. Skibsted 

Carotenoid scavenging of free radicals has been investigated in peroxidating 

methyl esters of unsaturated fatty acids using (i) metmyoglobin as a water-based 

free-radical initiator in a heterogeneous lipid/water system, and (ii) azo-bisisobutyronitrile 

as a free-radical initiator homogeneous chloroform solution. For 

the heterogeneous system, using a combination of electrochemical oxygen 

depletion measurements, spectrophotometric determination of lipid 

hydroperoxides and carotenoid degradation, it was demonstrated that each of the 

four carotenoids astaxanthin, β-carotene, canthaxanthin, and zeaxanthin protects 

the methyl esters against oxidation. The antioxidant effect increases with 

increasing carotenoid concentration increases with decreasing oxygen partial 

pressure (0.010 < 0.50 atm), and shows little dependence on the structure of the 

carotenoid. For a homogeneous solution, the effect of the structure of the 

carotenoid was further investigated, and it was shown that the stability of the four 

carotenoids in the oxidizing system are different, with the order of decreasing 

stability being: astaxanthin > canthaxanthin > β-carotene > zeaxanthin. Each of 

the four carotenoids can suppress lipid oxidation and the degree of suppression of 

peroxidation of methyl linoleate corresponds to the difference in stability. 

Antioxidant 

34

Pure & Appl. Chem., Vol. 63, No. 1, pp. 141-146, 1991. 

Printed in Great Britain. 

1991 IUPAC 

Biological functions and activities of animal carotenoids 

Wataru Miki 

Astaxanthin, on of the dominant carotenoids in marine animals, showed both a 

strong quenching effect against singlet oxygen, and a strong scavenging effect 

against free radicals. These effects are considered to be defense mechanisms in 

the animals for attacking these active oxygen species. The activities of 

astaxanthin are approximately 10 times stronger than those of other carotenoids 

that were tested, namely zeaxanthin, lutein, tunaxanthin, canthaxanthin and β- 

carotene, and 100 times greater than those of a tocopherol. Astaxanthin also 

showed strong activity as an inhibitor of lipid peroxidation mediated by these 

active forms of oxygen. From these results, astaxanthin has a the properties of a 

“SUPER VITAMIN E”. 

Antioxidant 

35

Am J Cardiol 2008;101[suppl]:20D–29D 

Biologic Activity of Carotenoids Related to Distinct Membrane 

Physicochemical Interactions 

Hyesun McNulty, PhD,a Robert F. Jacob, PhD,a and R. Preston Mason, 

PhDa,b,* 

Carotenoids are naturally occurring organic pigments that are believed to have 

therapeutic benefit in treating cardiovascular disease (CVD) because of their 

antioxidant properties. However, prospective randomized trials have failed to 

demonstrate a consistent benefit for the carotenoid _-carotene in patients at risk 

for CVD. The basis for this apparent paradox is not well understood but may be 

attributed to the distinct antioxidant properties of various carotenoids resulting 

from their structure-dependent physicochemical interactions with biologic 

membranes. To test this hypothesis, we measured the effects of astaxanthin, 

zeaxanthin, lutein, _-carotene, and lycopene on lipid peroxidation using model 

membranes enriched with polyunsaturated fatty acids. The correlative effects of 

these compounds on membrane structure were determined using small-angle x- 

ray diffraction approaches. The nonpolar carotenoids, lycopene and _-carotene, 

disordered the membrane bilayer and stimulated membrane lipid peroxidation 

(>85% increase in lipid hydroperoxide levels), whereas astaxanthin (a polar 

carotenoid) preserved membrane structure and exhibited significant antioxidant 

activity ( >40% decrease in lipid hydroperoxide levels). These results suggest that 

the antioxidant potential of carotenoids is dependent on their distinct membrane 

lipid interactions. This relation of structure and function may explain the 

differences in biologic activity reported for various carotenoids, with important 

therapeutic implications. 

rights reserved. 

Antioxidant 

36

Int. J. Vitam. Nutr. Res., 77(1), 2007, 3-11 

Effects pf Astaxanthin Supplementation on Lipid Peroxidation 

Jouni Karppi, Tiina H. Rissanen, kristiina Nyyssonen, Jari Kaikkonen, Anders G. 

Olsson, Sari Voutilainen and Jukka T. Salonen 

Abstract: Astaxanthin, the main carotenoid pigment in aquatic animals, has greater 

antioxidant activity in vitro (protecting against lipid peroxidation) and aa more 

polar configuration then other carotenoids. We investigated the effect of threemonth 

astaxanthin supplementation on lipid peroxidation in healthy non-smoking 

Finnish men, aged 19-33 years by using a randomized double-blind study design. 

Also absorption of astaxanthin from capsules into bloodstream and its safety were 

evaluate. The intervention group received two 4-mg astaxanthin (Astxan®) 

capsules daily, and the control group two identical-looking placebo capsules. 

Astaxanthin supplementation elevated plasma astaxanthin levels to 0.032 μmol/L 

(p

J Agric Food Chem. 2006 Mar 22;54(6):2418-23. 

Astaxanthin protects against oxidative stress and calcium-induced 

porcine lens protein degradation. 

Wu TH, Liao JH, Hou WC, Huang FY, Maher TJ, Hu CC. 

Department of Clinical Pharmacy, School of Pharmacy, Taipei Medical 

University, Taipei 110, Taiwan. thwu@tmu.edu.tw 

Astaxanthin (ASTX), a carotenoid with potent antioxidant properties, exists 

naturally in various plants, algae, and seafoods. In this study, we investigated the 

in vitro ability of ASTX to protect porcine lens crystallins from oxidative damage 

by iron-mediated hydroxyl radicals or by calcium ion-activated protease (calpain), 

in addition to the possible underlying biochemical mechanisms. ASTX (1 mM) 

was capable of protecting lens crystallins from being oxidized, as measured by 

changes in tryptophan fluorescence, in the presence of a Fenton reaction solution 

containing 0.2 mM Fe2+ and 2 mM H2O2. Sodium dodecyl sulfatepolyacrylamide 

gel electrophoresis analysis demonstrated that beta(high)- 

crystallin was the most vulnerable protein under these conditions of free radical 

exposure. The proteolysis of lens crystallins induced by calcium ion-activated 

calpain was also inhibited by ASTX (0.03-1 mM) as determined by daily 

measurement of the light-scattering intensity at 405 nm for five consecutive days. 

ASTX at 1 mM was as potent as a concentration of 0.1 mM calpain inhibitor E64 

in protecting the oxidative damage/hydrolysis of porcine crystallins. At a 

concentration of 1 mM, ASTX provided better protection than the endogenous 

antioxidant glutathione in terms of suppressing calcium-induced turbidity of lens 

proteins. Thin-layer chromatography analysis indicated that ASTX interacted with 

calcium ions to form complexes, which we believe interfere with the hydrolysis of 

lens crystallins by calcium-activated calpain. This in vitro study shows that ASTX 

is capable of protecting porcine lens proteins from oxidative insults and 

degradation by calcium-induced calpain. 

PMID: 16536628 [PubMed - indexed for MEDLIN 

Antioxidant 

38

Reprod Domest Anim. 2009 Nov 18. [Epub ahead of print] 

Antioxidative Effects of Astaxanthin against Nitric Oxide-Induced 

Oxidative Stress on Cell Viability and Gene Expression in Bovine Oviduct 

Epithelial Cell and the Developmental Competence of Bovine IVM/IVF 

Embryos. 

Jang HY, Ji SJ, Kim YH, Lee HY, Shin JS, Cheong HT, Kim JT, Park IC, Kong HS, Park 

CK, Yang BK. 

College of Animal Life Science, Kangwon National University, Chuncheon, Korea. 


Contents The aim of the present study was to elucidate the fundamental mechanism of 

bovine oviduct epithelial cell (BOEC) co-culture on developmental capacity of bovine in 

vitro oocyte maturation/in vitro fertilization (IVM/IVF) embryos. We examined the 

effects of astaxanthin against nitric oxide-induced oxidative stress on cell viability by 

MTT assay, lipid peroxidation (LPO) by using thiobarbituric acid (TBA) reaction for 

malondialdehyde (MDA) and the expression of antioxidant genes (CuZnSOD, MnSOD 

and Catalase) or apoptosis genes (Bcl-2, Caspase-3 and Bax) by RT-PCR in BOEC. We 

also evaluated the developmental rates of bovine IVM/IVF embryos co-cultured with 

BOEC pre-treated with astaxanthin (500 mum) in the presence or absence of sodium 

nitroprusside (SNP, 1000 mum) for 24 h. Cell viability in BOEC treated with SNP (50- 

2000 mum) lowered, while astaxanthin addition (50-500 mum) increased it in a dosedependent 

manner. Cell viability in astaxanthin plus SNP (1000 mum) gradually 

recovered according to the increase in astaxanthin additions (100-500 mm). The LPO in 

astaxanthin group (50-500 muM) gradually decreased in a dose dependent manner and 

among SNP or astaxanthin plus SNP group, SNP alone and astaxanthin (50 muM) plus 

SNP shown a significant increase than other groups (p < 0.05). Expression of apoptosis 

or antioxidant genes was detected by RT-PCR. Bcl-2 and antioxidant genes were detected 

in astaxanthin or astaxanthin plus SNP group, and Caspase-3 and Bax genes were only 

found in SNP group. When bovine IVM/IVF embryos were cultured for 6-7 days under 

co-culture system such as BOEC treated with astaxanthin in the presence or absence of 

SNP, the developmental ability to blastocysts in 500 mum astaxanthin group was the 

highest of all groups. These results suggest that astaxanthin has a antioxidative effect on 

cell viability and LPO of BOEC, and development of bovine IVM/IVF embryos due to 

the induction of antioxidant genes and suppression of apoptosis genes. 


39

Antioxidant 

Cell Biol Toxicol. 2010 Oct;26(5):457-67. Epub 2010 Mar 14. 

Astaxanthin prevents in vitro auto-oxidative injury in human 

lymphocytes. 

Bolin AP, Macedo RC, Marin DP, Barros MP, Otton R. 

Cellular Physiology Laboratory, Postgraduate Program-Health Science, CBS, Cruzeiro 

do Sul University, Tatuapé, São Paulo, Brazil. 


Upon mitogen sensitization, lymphocytes undergo proliferation by oxyradical-based 

mechanisms. Through continuous resting-restimulation cycles, lymphocytes accumulate 

auto-induced oxidative lesions which lead to cell dysfunction and limit their viability. 

Astaxanthin (ASTA) is a nutritional carotenoid that shows notable antioxidant properties. 

This study aims to evaluate whether the in vitro ASTA treatment can limit oxyradical 

production and auto-oxidative injury in human lymphocytes. Activated lymphocytes 

treated with 5 microM ASTA showed immediate lower rates of O(2)(*-) /H(2)O(2) 

production whilst NO* and intracellular Ca(2+) levels were concomitantly enhanced 

(24 h), the cytotoxicity test for ASTA showed a 

sigmoidal dose-response curve (LC50 = 11.67 +/- 0.42 microM), whereas higher 

activities of superoxide dismutase and catalase in 5 microM ASTA-treated lymphocytes 

were associated to significant lower indexes of oxidative injury. On the other hand, lower 

proliferative scores of ASTA lymphocytes might be a result of diminished intracellular 

levels of pivotal redox signaling molecules, such as H(2)O(2). Further studies are 

necessary to establish the ASTA-dose compensation point between minimizing oxidative 

damages and allowing efficient redox-mediated immune functions, such as proliferation, 

adhesion, and oxidative burst. 


40

Antioxidant 

Eur J Nutr. 2010 Apr 2. [Epub ahead of print] 

Astaxanthin addition improves human neutrophils 

function: in vitro study. 

Macedo RC, Bolin AP, Marin DP, Otton R. 

Postgraduate Program, Health Science, CBS, Cruzeiro do Sul University, Avenida 

Regente Feijó, 1295. Tatuapé, São Paulo, SP, CEP 03342-000, Brazil. 


PURPOSE: The aim of the present study was to evaluate the in vitro effect of carotenoid 

astaxanthin (ASTA) on the phagocytic and microbicidal capacities, cytokine release, and 

reactive oxygen species production in human neutrophils. 

METHODS: The following parameters were evaluated: cytotoxic effect of ASTA on 

human neutrophils viability, phagocytic and microbicidal capacities of neutrophils by 

using Candida albicans assay, intracellular calcium mobilization (Fura 2-AM fluorescent 

probe), superoxide anion (lucigenin and DHE probes), hydrogen peroxide (H(2)O(2), 

phenol red), and nitric oxide (NO.) (Griess reagent) production, activities of antioxidant 

enzymes (total/Mn-SOD, CAT, GPx, and GR), oxidative damages in biomolecules 

(TBARS assay and carbonyl groups), and cytokine (IL-6 and TNF-alpha) release. 

RESULTS: Astaxanthin significantly improves neutrophil phagocytic and microbicidal 

capacity, and increases the intracellular calcium concentration and NO. production. Both 

functional parameters were accompanied by a decrease in superoxide anion and hydrogen 

peroxide and IL-6 and TNF-alpha production. Oxidative damages in lipids and proteins 

were significantly decreased after ASTA-treatment. 

CONCLUSIONS: Taken together our results are supportive to a beneficial effect of 

astaxanthin-treatment on human neutrophils function as demonstrated by increased 

phagocytic and fungicide capacity as well as by the reduced superoxide anion and 

hydrogen peroxide production, however, without affecting neutrophils capacity to kill C. 

albicans. This process appears to be mediated by calcium released from intracellular 

storages as well as nitric oxide production. 


Antioxidant 

41

Phytother Res. 2010 Jan;24(1):54-9. 

Cytoprotective role of astaxanthin against glycated 

protein/iron chelate-induced toxicity in human 

umbilical vein endothelial cells. 

Nishigaki I, Rajendran P, Venugopal R, Ekambaram G, Sakthisekaran D, Nishigaki Y. 

NPO International Laboratory of Biochemistry, 1-166 Uchide, Nakagawa-ku Nagoya 

454-0926, Japan. nishigaki@se.starcat.ne.jp 






interfering with ROS generation in these cells. Glycated fetal bovine serum (GFBS) was 

prepared by incubating fetal bovine serum (FBS) with high-concentration glucose. 

Stimulation of cultured HUVECs with 50 mm 1 mL of GFBS significantly enhanced 

lipid peroxidation and decreased antioxidant enzyme activities and levels of phase II 

enzymes. However, preincubation of the cultures with ASX resulted in a marked decrease 

in the level of lipid peroxide (LPO) and an increase in the levels of antioxidant enzymes 

in an ASX concentration-dependent manner. These results demonstrate that ASX could 

inhibit LPO formation and enhance the antioxidant enzyme status in GFBS/iron chelateexposed 

endothelial cells by suppressing ROS generation, thereby limiting the effects of 

the AGE-RAGE interaction. The results indicate that ASX could have a beneficial role 

against glycated protein/iron chelate-induced toxicity by preventing lipid and protein 

oxidation and increasing the activity of antioxidant enzymes. 


Antioxidant 

42

Plant Foods Hum Nutr. 2011 Oct 1. [Epub ahead of print] 

Positive Effects of Astaxanthin on Lipid 

Profiles and Oxidative Stress in 

Overweight Subjects. 

Choi HD, Youn YK, Shin WG. 

Source 

College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National 

University, San 56-1, Sillim-Dong, Gwanak-Gu, Seoul, 151-742, South Korea. 


Astaxanthin, a carotenoid, has antioxidant activity as well as many positive effects, such 

as anticancer and anti-inflammatory effects. We performed a randomized, double-blind, 

placebo-controlled study to investigate the effects of astaxanthin on lipid profiles and 

oxidative stress in overweight and obese adults in Korea. In total, 27 subjects with body 

mass index >25.0 kg/m(2) were enrolled and randomly assigned into two groups 

administered astaxanthin or placebo capsules for 12 weeks. Total cholesterol, 

triglycerides, high density lipoprotein (HDL) cholesterol, low density lipoprotein (LDL) 

cholesterol, apolipoprotein A1 (ApoA1), and apolipoprotein B (ApoB) were measured 

before and after intervention. Malondialdehyde (MDA), isoprostane (ISP), superoxide 

dismutase (SOD), and total antioxidant capacity (TAC), as oxidative stress biomarkers, 

were measured at baseline and at 4, 8, and 12 weeks after intervention. LDL cholesterol 

and ApoB were significantly lower after treatment with astaxanthin, compared with the 

start of administration, whereas none of the lipid profiles was changed in the placebo 

group. At the baseline, all four biomarkers were not significantly different between the 

two groups. Compared with the placebo group, MDA and ISP were significantly lower, 

but TAC was significantly higher in the astaxanthin group at 12 weeks. These results 

suggest that supplementary astaxanthin has positive effects by improving the LDL 

cholesterol, ApoB, and oxidative stress biomarkers. 


Antioxidant 

43

Phytother Res. 2011 Apr 8. doi: 10.1002/ptr.3494. [Epub ahead of print] 

Effects of Astaxanthin on Oxidative Stress 

in Overweight and Obese Adults. 

Choi HD, Kim JH, Chang MJ, Kyu-Youn Y, Shin WG. 

Source 

College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National 

University, San 56-1, Sillim-Dong, Gwanak-Gu, Seoul, 151-742, South Korea. 


Oxidative stress is caused by an imbalance between the antioxidant and the reactive 

oxygen species, which results in damage to cells or tissues. Recent studies have reported 

that oxidative stress is involved in obesity, in addition to many other human diseases and 

aging. A prospective, randomized, double-blind study was performed to investigate the 

effect of astaxanthin (ASX), which is known to be a potent antioxidant, on oxidative 

stress in overweight and obese adults in Korea. Twenty-three adults with 

BMI > 25.0 kg/m(2) enrolled in this study and were randomly assigned to two dose 

groups: ASX 5 mg and 20 mg once daily for 3 weeks. Malondialdehyde (MDA), 

isoprostane (ISP), superoxide dismutase (SOD) and total antioxidant capacity (TAC), as 

oxidative stress biomarkers, were measured at baseline and 1, 2 and 3 weeks after ASX 

administration. Compared with baseline, the MDA (by 34.6% and 35.2%) and ISP (by 

64.9% and 64.7%) levels were significantly lowered, whereas SOD (by 193% and 194%) 

and TAC (by 121% and 125%) levels were significantly increased in two dose groups 

after the 3 week intervention. This study revealed that supplemental ASX for 3 weeks 

improved oxidative stress biomarkers by suppressing lipid peroxidation and stimulating 

the activity of the antioxidant defense system. Copyright © 2011 John Wiley & Sons, 

Ltd. 

Copyright © 2011 John Wiley & Sons, Ltd. 


Antioxidant 

44

Can J Physiol Pharmacol. 2010 Oct;88(10):977-85. 

Retinol-deficient rats can convert a 

pharmacological dose of astaxanthin to retinol: 

antioxidant potential of astaxanthin, lutein, and β- 

carotene. 

Sangeetha RK, Baskaran V. 

Source 

Department of Biochemistry and Nutrition, Central Food Technological Research 

Institute, CSIR, Mysore, Karnataka, India. 


Retinol (ROH) and provitamin-A carotenoids are recommended to treat ROH deficiency. 

Xanthophyll carotenoids, being potent antioxidants, can modulate health disorders. We 

hypothesize that nonprovitamin-A carotenoids may yield ROH and suppress lipid 

peroxidation under ROH deficiency. This study aimed to (i) study the possible 

bioconversion of astaxanthin and lutein to ROH similar to β-carotene and (ii) determine 

the antioxidant potential of these carotenoids with reference to Na(+)/K(+)-ATPase, 

antioxidant molecules, and lipid peroxidation (Lpx) induced by ROH deficiency in rats. 

ROH deficiency was induced in rats (n = 5 per group) by feeding a diet devoid of ROH. 

Retinol-deficient (RD) rats were gavaged with astaxanthin, lutein, β-carotene, or peanut 

oil alone (RD group) for 7 days. Results show that the RD group had lowered plasma 

ROH levels (0.3 µmol/L), whereas ROH rose in astaxanthin and β-carotene groups (4.9 

and 5.7 µmol/L, respectively), which was supported by enhanced (69% and 70%) 

intestinal β-carotene 15,15'-monooxygenase activity. Astaxanthin, lutein, and β-carotene 

lowered Lpx by 45%, 41%, and 40% (plasma), respectively, and 59%, 64%, and 60% 

(liver), respectively, compared with the RD group. Lowered Na(+)/K(+)-ATPase and 

enhanced superoxide dismutase, catalase, and glutathione-S-transferase activities support 

the lowered Lpx. To conclude, this report confirms that astaxanthin is converted into β- 

carotene and ROH in ROH-deficient rats, and the antioxidant potential of carotenoids 

was in the order astaxanthin > lutein > β-carotene. 


Antioxidant 

45

Fast regeneration of carotenoids from radical 

cations by isoflavonoid dianions: importance of 

the carotenoid keto group for electron transfer. 

Han RM, Chen CH, Tian YX, Zhang JP, Skibsted LH. 

Source 

Department of Chemistry, Renmin University of China, Beijing 100872, People's 

Republic of China. 


Electron transfer to radical cations of beta-carotene, zeaxanthin, canthaxanthin, and 

astaxanthin from each of the three acid/base forms of the diphenolic isoflavonoid 

daidzein and its C-glycoside puerarin, as studied by laser flash photolysis in 

homogeneous methanol/chloroform (1/9) solution, was found to depend on carotenoid 

structures and more significantly on the deprotonation degree of the isoflavonoids. None 

of the carotenoid radical cations reacted with the neutral forms of the isoflavonoids while 

the monoanionic and dianionic forms of the isoflavonoids regenerated the oxidized 

carotenoid. Electron transfer to the beta-carotene radical cation from the puerarin dianion 

followed second order kinetics with the rate constant at 25 degrees C k(2) = 5.5 x 10(9) 

M(-1) s(-1), zeaxanthin 8.5 x 10(9) M(-1) s(-1), canthaxanthin 6.5 x 10(10) M(-1) s(-1), 

and astaxanthin 11.1 x 10(10) M(-1) s(-1) approaching the diffusion limit and 

establishing a linear free energy relationship between rate constants and driving force. 

Comparable results found for the daidzein dianion indicate that the steric hindrance from 

the glucoside is not important suggesting the more reducing but less acidic 4'-OH/4'-O(-) 

as electron donors. On the basis of the rate constants obtained from kinetic analyses, the 

keto group of carotenoids is concluded to facilitate electron transfer. The driving force 

was estimated from oxidation potentials, as determined by cyclic-voltametry for puerarin 

and daidzein in aqueous solutions at varying pH conditions, which led to the standard 

reduction potentials E degrees = 1.13 and 1.10 V versus NHE corresponding to the 

uncharged puerarin and daidzein. For pH > pK(a2), the apparent potentials of both 

puerarin and daidzein became constants and were E degrees = 0.69 and 0.65 V, 

respectively. Electron transfer from isoflavonoids to the carotenoid radical cation, as 

formed during oxidative stress, is faster for astaxanthin than for the other carotenoids, 

which may relate to astaxanthins more effective antioxidative properties and in 

agreement with the highest electron accepting index of astaxanthin. 


Antioxidant 

46

J Clin Biochem Nutr. 2010 Sep;47(2):130-7. Epub 2010 Jun 22. 

Evaluation of therapeutic effects of astaxanthin on 

impairments in salivary secretion. 

Yamada T, Ryo K, Tai Y, Tamaki Y, Inoue H, Mishima K, Tsubota K, Saito I. 

Source 

Department of Pathology, Tsurumi University School of Dental Medicine, 2-1-3, 

Tsurumi, Tsurumi-ku, Yokohama 230-8501, Japan. 


The involvement of reactive oxygen species (ROS) in the pathophysiology of Sjögren's 

syndrome (SS), an autoimmune disorder, and irradiation-induced impairments in salivary 

secretion has been reported. Meanwhile, the strong antioxidant astaxanthin (Ast) has been 

suggested to have therapeutic effects on various diseases. In the present study, we 

examined the ROS scavenging capacity of Ast using a human salivary gland epithelial 

cell line (HSY) and investigated the effects of Ast on salivary secretion in a mouse model 

of irradiation-induced salivary gland dysfunction. Furthermore, we performed a clinical 

study of Ast in six SS patients and six normal individuals, quantifying the volume of 

saliva secretion and the level of oxidative stress markers in the saliva. Ast partially 

suppressed hydrogen peroxide-induced ROS in HSY cells. The mouse model 

demonstrated that the pre-administration of Ast resulted in the suppression of irradiationinduced 

hyposalivation. Furthermore, the administration of Ast appeared to increase 

salivary output in both the SS and normal groups. The level of oxidative stress marker, 

hexanoyl-lysine, in the saliva was reduced after Ast intake. These results suggest that Ast 

might act as an ROS scavenger, providing benefits to SS patients with impaired salivary 

secretion. 

PMID: 20838568 [PubMed] 

PMCID: PMC2935153 

Antioxidant 

47

J Med Food. 2011 Sep 1. [Epub ahead of print] 

Protective Effects of Haematococcus Astaxanthin on Oxidative Stress in 

Healthy Smokers. 

Kim JH, Chang MJ, Choi HD, Youn YK, Kim JT, Oh JM, Shin WG. 

Source 

1 College of Pharmacy, Clinical Pharmaceutical Education and Research Institute, Seoul 

National University , Seoul, Korea. 


Abstract Free radicals induced by cigarette smoking have been strongly linked to 

increased oxidative stress in vivo, contributing to the pathobiology of various diseases. 

This study was performed to investigate the effects of Haematococcus astaxanthin 

(ASX), which has been known to be a potent antioxidant, on oxidative stress in smokers. 

Thirty-nine heavy smokers (≥20 cigarettes/day) and 39 non-smokers were enrolled in this 

study. Smokers were randomly divided into three dosage groups to receive ASX at doses 

of 5, 20, or 40 mg (n=13, each) once daily for 3 weeks. Oxidative stress biomarkers such 

as malondialdehyde, isoprostane, superoxide dismutase, and total antioxidant capacity, 

and ASX levels in plasma were measured at baseline and after 1, 2, and 3 weeks of 

treatment. Compared with baseline, the plasma malondialdehyde and isoprostane levels 

decreased, whereas superoxide dismutase level and total antioxidant capacity increased in 

all ASX intervention groups over the 3-week period. In particular, isoprostane levels 

showed a significant dose-dependent decrease after ASX intake. The results suggest that 

ASX supplementation might prevent oxidative damage in smokers by suppressing lipid 

peroxidation and stimulating the activity of the antioxidant system in smokers. 


Antioxidant 

48

Eur J Nutr. 2011 Oct 5. [Epub ahead of print] 

Combined fish oil and astaxanthin supplementation modulates 

rat lymphocyte function. 

Otton R, Marin DP, Bolin AP, de Cássia Santos Macedo R, Campoio TR, Fineto C Jr, 

Guerra BA, Leite JR, Barros MP, Mattei R. 

Source 

Postgraduate Program, Health Sciences, CBS, Cruzeiro do Sul University, Av. Regente 

Feijó, 1295, Sao Paulo, SP, 03342000, Brazil, rosemari.otton@cruzeirodosul.edu.br. 


PURPOSE: Higher intakes of n-3 polyunsaturated fatty acids that are abundant in marine 

fishes have been long described as a "good nutritional intervention" with increasing 

clinical benefits to cardiovascular health, inflammation, mental, and neurodegenerative 

diseases. The present study was designed to investigate the effect of daily fish oil (FO- 

10 mg EPA/kg body weight (BW) and 7 mg DHA/kg BW) intake by oral gavage 

associated with the antioxidant astaxanthin (ASTA-1 mg/kg BW) on the redox 

metabolism and the functional properties of lymphocytes from rat lymph nodes. 

METHODS: This study was conducted by measurements of lymphocyte proliferation 

capacity, ROS production [superoxide (O (2) (•-) ) and hydrogen peroxide (H(2)O(2))], 

nitric oxide (NO(•)) generation, intracellular calcium release, oxidative damage to lipids 

and proteins, activities of major antioxidant enzymes, GSH/GSSG content, and cytokines 

release. 

RESULTS: After 45 days of FO + ASTA supplementation, the proliferation capacity of 

activated T- and B-lymphocytes was significantly diminished followed by lower levels of 

O (2) (•-) , H(2)O(2) and NO(•) production, and increased activities of total/SOD, GR 

and GPx, and calcium release in cytosol. ASTA was able to prevent oxidative 

modification in cell structures through the suppression of the oxidative stress condition 

imposed by FO. L: -selectin was increased by FO, and IL-1β was decreased only by 

ASTA supplementation. 

CONCLUSION: We can propose that association of ASTA with FO could be a good 

strategy to prevent oxidative stress induced by polyunsaturated fatty acids and also to 

potentiate immuno-modulatory effects of FO. 


Antioxidant 

49

Toxicol In Vitro. 2011 Oct;25(7):1448-56. Epub 2011 Apr 27. 

Oxidative stress in human lymphocytes 

treated with fatty acid mixture: role of 

carotenoid astaxanthin. 

Campoio TR, Oliveira FA, Otton R. 

Source 

Postgraduate Program - Health Sciences - CBS, Cruzeiro do Sul University, 03342000 

Sao Paulo, SP, Brazil. 


Fatty acids (FA) have been shown to alter leukocyte function, and depending on 

concentration and type, they can modulate both inflammatory and immune responses. 

Astaxanthin (ASTA) is a carotenoid that shows notable antioxidant properties. In the 

present study we propose to evaluate the oxidative stress in human lymphocytes induced 

by a FA mixture and the possible protective role of ASTA. The present study showed that 

the FA mixture at 0.3mM caused a marked increase in the production of superoxide 

anion, hydrogen peroxide and nitric oxide, which was accompanied by an increase in 

total-SOD activity, in TBARS levels and a reduction of catalase activity and content of 

GSH and free thiol groups. The FA mixture also promoted an increase in intracellular 

Ca(2+) mobilization and in the proliferative capacity of B-lymphocytes. The addition of 

ASTA (2 μM) partially decreased the ROS production and TBARS levels and increased 

the levels of free thiol groups. ASTA decreased the proliferative capacity of cells treated 

with FA but not by reducing intracellular calcium concentration. Based on these results 

we can conclude that ASTA can partially prevent oxidative stress in human lymphocytes 

induced by a fatty acid mixture, probably by blenching/quenching free radical 

production. 

Copyright © 2011 Elsevier Ltd. All rights reserved. 


Antioxidant 

50

J Nutr. 2011 Sep;141(9):1611-7. Epub 2011 Jul 6. 

Astaxanthin-rich extract from the green alga Haematococcus 

pluvialis lowers plasma lipid concentrations and enhances 

antioxidant defense in apolipoprotein E knockout mice. 

Yang Y, Seo JM, Nguyen A, Pham TX, Park HJ, Park Y, Kim B, Bruno RS, Lee J. 

Source 

Department of Nutritional Sciences, University of Connecticut, Storrs, CT, USA. 


Dyslipidemia and oxidative stress contribute to atherogenesis. Astaxanthin (ASTX) is a 

red-colored carotenoid well known for its high antioxidant capacity. However, its effects 

on lipid metabolism and antioxidant defense mechanisms have received only limited 

investigation. We fed male apoE knockout (apoE)(-/-) mice, a mouse model for 

atherosclerosis, a high-fat (15%)/high-cholesterol (0.2%) diet alone (control) or 

supplemented with ASTX-rich Hematococcus pluvialis extract (0.03% ASTX by weight) 

for 4 wk. ASTX-fed apoE(-/-) mice had significantly lower plasma total cholesterol and 

TG concentrations than controls, but body weight and plasma alanine aminotransferase 

and aspartate aminotransferase did not differ between the groups. qRT-PCR analysis 

demonstrated significantly greater mRNA levels of LDL receptor (LDLR), 3-hydroxy-3- 

methylglutaryl CoA reductase, and sterol regulatory element binding protein 2 (SREBP- 

2) and greater mature SREBP-2 protein in the livers of ASTX-fed mice, indicating that 

increased LDLR expression may be responsible for the hypocholesterolemic effect of 

ASTX. Hepatic lipogenic gene expression was not altered, but carnitine palmitoyl 

transferase 1, acetyl-CoA carboxylase β, and acyl-CoA oxidase mRNA abundance were 

significantly increased by ASTX supplementation, suggesting the TG-lowering effect of 

ASTX may be due to increased fatty acid β-oxidation in the liver. Expression of the 

nuclear factor E2 related factor 2-responsive endogenous antioxidant gene also was 

induced with concomitantly lower glutathione disulfide levels in the livers of ASTX-fed 

apoE(-/-) mice compared to controls. In conclusion, these results suggest that 

supplementation of ASTX-rich H. pluvialis extract improves cholesterol and lipid 

metabolism as well as antioxidant defense mechanisms, all of which could help mitigate 

the progression of atherosclerosis. 

PMID:21734060 [PubMed - in process] 

Antioxidant 

51

Fish Physiol Biochem. 2011 Jun 22. [Epub ahead of print] 

Effects of Haematococcus pluvialis 

supplementation on antioxidant system 

and metabolism in rainbow trout 

(Oncorhynchus mykiss). 

Sheikhzadeh N, Tayefi-Nasrabadi H, Khani Oushani A, Najafi Enferadi MH. 

Source 

Department of Food Hygiene and Aquatic Animals, Faculty of Veterinary Medicine, 

University of Tabriz, Tabriz, Iran, nsheikh@tabrizu.ac.ir. 


Effects of commercial source for astaxanthin (Haematococcus pluvialis) (H.p) on 

antioxidant power, specific marker enzymes, and some metabolites were examined in 

rainbow trout (Oncorhynchus mykiss). Fish were fed on diets containing 1, 3, and 10 g 

microalga kg(-1) feed for 30 days. Serum total antioxidant activity and lipid peroxidation 

product, indicated by malondialdehyde (MDA), significantly enhanced with different 

doses of administration, indicating the elevated antioxidant status in all treatment groups. 

In group fed with high dose of alga, significantly elevated aspartate aminotransferase 

activity (AST) was noted, indicating damage of normal liver function in this group. 

Alkaline phosphatase (ALP) and alanine aminotransferase (ALT) were not affected in all 

groups. Although serum total protein remained unaffected, serum glucose level was 

decreased significantly in lower doses of administration. Furthermore, triglyceride and 

cholesterol levels showed significant decrease in 3 g kg(-1) microalga group by 

modulation of lipid metabolism in this group. On the other hand, in highest dose, 

significant increase in lipids was observed, indicating the slight dysfunction in lipid 

metabolism in this treatment group. The present study suggests that Haematococcus 

pluvialis especially in dose of 3 g kg(-1) feed administration may effectively enhance the 

antioxidant system and some biochemical parameters in rainbow trout. 


Antioxidant 

52

Yao Xue Xue Bao. 2011 May;46(5):521-6. 

[Astaxanthin inhibits sodium azide-induced cytotoxicity in 

hepatocyte L-02 cells probably by H+ transferring function]. 


Ma J, Chen HM, Yan XJ, Wang F, Xu WF. 

Source 

Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, 

China. 


This study is to investigate the protective effect of astaxanthin against injured hepatocyte 

L-02 cells induced by sodium azide (NaN3) and reveal the possible mechanisms. 

Hepatocyte L-02 cells were exposed to 100 mmol.L-1 NaN3 with various concentrations 

of astaxanthin pre-incubated, then the cell viability was measured by MTT method; The 

level of reactive oxygen species (ROS) was determined by DCFH-DA method; The 

changes of mitochondrial membrane potential (MMP) and apoptosis ratio were detected 

by JC-1 method and Annexin V-FITC/PI double stain method, respectively. Results 

showed that after cells were exposed to 100 mmol.L-1 NaN3 for 3 hours, the cell 

viability significantly decreased; ROS level and the percentage of late phase apoptosis 

increased obviously; MMP was also declined. When cells were pretreated with 

astaxanthin, the cell damage and late phase apoptosis ratio reduced and MMP was 

maintained. However, the level of ROS showed insignificant decrease (P>0.05). The 

beneficial concentration of astaxanthin in improving cell viability and MMP was not in a 

dose dependent manner and the most effective of which was 0.10 nmol.L-1 (P

Reprod Domest Anim. 2010 Dec;45(6):e387-91. doi: 10.1111/j.1439-0531.2010.01584.x. 

Effects of astaxanthin-containing oil on development and stress-related 

gene expression of bovine embryos exposed to heat stress. 

Namekawa T, Ikeda S, Sugimoto M, Kume S. 

Source 

Laboratory of Animal Physiology and Functional Anatomy, Graduate School of 

Agriculture, Kyoto University, Kyoto, Japan. 


Early bovine embryos are vulnerable to heat stress during the first few days after 

fertilization. The inhibitory effect of heat stress on embryonic development is known to 

be associated with oxidative stress, which can be attenuated by antioxidants. In the 

present study, we focused on the use of astaxanthin as an antioxidant and examined the 

effects of astaxanthin-containing oil (Ax) on post-fertilization development of bovine 

embryos subjected to heat stress in vitro and the expression of stress-related genes. 

Bovine 1-cell embryos were in vitro produced by in vitro maturation and fertilization 

(IVF) of oocytes recovered from abattoir-derived ovaries. At 20 h post-insemination 

(hpi, 0 h = the start of IVF), the embryos were introduced in modified synthetic 

oviduct fluid supplemented with 25 ppm of Ax (concentration of astaxanthin was 0.25 

ppm) or vehicle (dimethyl sulfoxide) up to 72 hpi. The embryos were basically cultured 

at 38.5°C, and in the heat stress group, embryos were exposed twice to 40.5°C for 10 h 

(at 20-30 and 44-54 hpi). Under the condition without the Ax treatment, the cleavage 

rate, rate of development to the 5-8 cell stage, blastocyst yield from cultured embryos and 

that from cleaved embryos were lower in the heat stress group than in the group not 

subjected to heat stress (p < 0.05). In the heat stress group, the rate of development to 

the 5-8 cell stage was improved (p < 0.05) by the addition of Ax. Subsequently, we 

performed semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) 

to investigate the effects of heat stress and Ax on the mRNA expression of Src homology 

2 domain-containing transforming protein C1 (SHC1), an oxidative stress adaptor 

protein, and superoxide dismutase 2 (SOD2), a mitochondrial reactive oxygen species 

(ROS) scavenger. In 5-8 cell embryos at 72 hpi, the mRNA expression levels of SHC1 

and SOD2 were lower in the Ax- and heat-treated group than in the other groups (p < 

0.05). These results suggest that Ax added to the culture medium ameliorates the 

embryonic development impaired by heat stress with its altering effects on the expression 

of stress-related genes. 

© 2010 Blackwell Verlag GmbH. 


Antioxidant 

54

Reprod Domest Anim. 2010 Dec;45(6):967-74. doi: 10.1111/j.1439-0531.2009.01469.x. 

Antioxidative effects of astaxanthin against nitric oxide-induced 

oxidative stress on cell viability and gene expression in bovine oviduct 

epithelial cell and the developmental competence of bovine IVM/IVF 

embryos. 

Jang HY, Ji SJ, Kim YH, Lee HY, Shin JS, Cheong HT, Kim JT, Park IC, Kong HS, Park CK, 

Yang BK. 

Source 

College of Animal Life Science, Kangwon National University, Chuncheon, Korea. 


The aim of the present study was to elucidate the fundamental mechanism of bovine 

oviduct epithelial cell (BOEC) co-culture on developmental capacity of bovine in vitro 

oocyte maturation/in vitro fertilization (IVM/IVF) embryos. We examined the effects of 

astaxanthin against nitric oxide-induced oxidative stress on cell viability by MTT assay, 

lipid peroxidation (LPO) by using thiobarbituric acid (TBA) reaction for 

malondialdehyde (MDA) and the expression of antioxidant genes (CuZnSOD, MnSOD 

and Catalase) or apoptosis genes (Bcl-2, Caspase-3 and Bax) by RT-PCR in BOEC. We 

also evaluated the developmental rates of bovine IVM/IVF embryos co-cultured with 

BOEC pre-treated with astaxanthin (500 μM) in the presence or absence of sodium 

nitroprusside (SNP, 1000 μM) for 24 h. Cell viability in BOEC treated with SNP (50- 

2000 μM) lowered, while astaxanthin addition (50-500 μM) increased it in a dosedependent 

manner. Cell viability in astaxanthin plus SNP (1000 μM) gradually 

recovered according to the increase in astaxanthin additions (100-500 mM). The LPO 

in astaxanthin group (50-500 μM) gradually decreased in a dose dependent manner and 

among SNP or astaxanthin plus SNP group, SNP alone and astaxanthin (50 μM) plus 

SNP shown a significant increase than other groups (p < 0.05). Expression of 

apoptosis or antioxidant genes was detected by RT-PCR. Bcl-2 and antioxidant genes 

were detected in astaxanthin or astaxanthin plus SNP group, and Caspase-3 and Bax 

genes were only found in SNP group. When bovine IVM/IVF embryos were cultured for 

6-7 days under co-culture system such as BOEC treated with astaxanthin in the 

presence or absence of SNP, the developmental ability to blastocysts in 500 μM 

astaxanthin group was the highest of all groups. These results suggest that astaxanthin has 

a antioxidative effect on cell viability and LPO of BOEC, and development of bovine 

IVM/IVF embryos due to the induction of antioxidant genes and suppression of apoptosis 

genes. 

© 2009 Blackwell Verlag GmbH. 


Antioxidant 

55

Photosynth Res. 2010 Nov;106(1-2):155-77. Epub 2010 Aug 13. 

Secondary ketocarotenoid astaxanthin biosynthesis in 

algae: a multifunctional response to stress. 

Lemoine Y, Schoefs B. 

Source 

University Lille Nord de France, UMR 8187 LOG CNRS/University Lille 1, Bât SN2, 

59655 Villeneuve d'Ascq Cedex, France. 


Under stressful environments, many green algae such as Haematococcus pluvialis 

accumulate secondary ketocarotenoids such as canthaxanthin and astaxanthin. The 

carotenogenesis, responsible for natural phenomena such as red snows, generally 

accompanies larger metabolic changes as well as morphological modifications, i.e., the 

conversion of the green flagellated macrozoids into large red cysts. Astaxanthin 

accumulation constitutes a convenient way to store energy and carbon, which will be 

used for further synthesis under less stressful conditions. Besides this, the presence of 

high amount of astaxanthin enhances the cell resistance to oxidative stress generated by 

unfavorable environmental conditions including excess light, UV-B irradiation, and 

nutrition stress and, therefore, confers a higher survival capacity to the cells. This better 

resistance results from the quenching of oxygen atoms for the synthesis itself as well as 

from the antioxidant properties of the astaxanthin molecules. Therefore, astaxanthin 

synthesis corresponds to a multifunctional response to stress. In this contribution, the 

various biochemical, genetic, and molecular data related to the biosynthesis of 

ketocarotenoids by Haematococcus pluvialis and other taxa are reviewed and compared. 

A tentative regulatory model of the biochemical network driving astaxanthin production 

is proposed. 


Antioxidant 

56

Br J Nutr. 2010 Oct;104(7):980-8. Epub 2010 Jun 14. 

Effects of dietary lipid, vitamins and minerals on 

total amounts and redox status of glutathione and 

ubiquinone in tissues of Atlantic salmon (Salmo 

salar): a multivariate approach. 

Hamre K, Torstensen BE, Maage A, Waagbø R, Berge RK, Albrektsen S. 

Source 

National Institute of Nutrition and Seafood Research, PO Box 176, Sentrum, N-5804 

Bergen, Norway. kristin.hamre@nifes.no 


The hypothesis of the present study was that Atlantic salmon (Salmo salar) would 

respond to large variations in supplementation of dietary pro- and antioxidants, and 

marine lipid, with adjustment of the endogenously synthesised antioxidants, glutathione 

(GSH) and ubiquinone (UQ). An experiment with 2(7-3) reduced factorial design (the 

number of cases reduced systematically from 2(7) (full design) to 2(4) (reduced design)) 

was conducted, where vitamins, minerals and lipid were supplemented in the diet at high 

and low levels. For the vitamins and minerals the high levels were chosen to be just 

below anticipated toxic levels and the low levels were just above the requirement 

(vitamin C, 30 and 1000 mg/kg; vitamin E, 70 and 430 mg/kg; Fe, 70 and 1200 mg/kg; 

Cu, 8 and 110 mg/kg; Mn, 12 and 200 mg/kg). For astaxanthin, the dietary levels were 10 

and 50 mg/kg and for lipid, 150 and 330 g/kg. The experiment was started with postsmolts 

(148 (sd 17 g)) and lasted for 5 months. The only effect on GSH was a minor 

increase ( < 10 %) in total concentration in the liver in response to high dietary lipid. 

GSH redox state was not affected. UQ responded to dietary lipid, astaxanthin and vitamin 

E, both with regard to total concentration and redox state. Except for an effect of Fe on 

plasma GSH, the trace elements and vitamin C had no effect on tissue levels and 

oxidation state of GSH and UQ. This shows that the endogenous redox state is quite 

robust with regard to variation of dietary pro- and antioxidants in Atlantic salmon. 


Antioxidant 

57

J Nutr Biochem. 2010 May;21(5):381-9. Epub 2009 May 7. 

Astaxanthin protects mitochondrial redox state and functional 

integrity against oxidative stress. 

Wolf AM, Asoh S, Hiranuma H, Ohsawa I, Iio K, Satou A, Ishikura M, Ohta S. 

Source 

Department of Biochemistry and Cell Biology, Institute of Development and Aging 

Sciences, Nippon Medical School, Nakahara-ku, Kawasaki, Kanagawa, Japan. 


Mitochondria combine the production of energy with an efficient chain of reductionoxidation 

(redox) reactions but also with the unavoidable production of reactive oxygen 

species. Oxidative stress leading to mitochondrial dysfunction is a critical factor in many 

diseases, such as cancer and neurodegenerative and lifestyle-related diseases. Effective 

antioxidants thus offer great therapeutic and preventive promise. Investigating the 

efficacy of antioxidants, we found that a carotenoid, astaxanthin (AX), decreased 

physiologically occurring oxidative stress and protected cultured cells against strong 

oxidative stress induced with a respiratory inhibitor. Moreover, AX improved 

maintenance of a high mitochondrial membrane potential and stimulated respiration. 

Investigating how AX stimulates and interacts with mitochondria, a redox-sensitive 

fluorescent protein (roGFP1) was stably expressed in the cytosol and mitochondrial 

matrix to measure the redox state in the respective compartments. AX at nanomolar 


Additionally, AX improved the ability of mitochondria to remain in a reduced state under 

oxidative challenge. Taken together, these results suggest that AX is effective in 

improving mitochondrial function through retaining mitochondria in the reduced state. 

Copyright 2010 Elsevier Inc. All rights reserved. 


Antioxidant 

58

Mutat Res. 2010 Feb;696(1):69-80. Epub 2009 Dec 28. 

Astaxanthin intervention ameliorates cyclophosphamideinduced 

oxidative stress, DNA damage and early 

hepatocarcinogenesis in rat: role of Nrf2, p53, p38 and phase- 

II enzymes. 


Source 

Department of Pharmacology and Toxicology, National Institute of Pharmaceutical 

Education and Research (NIPER), Sector-67, S.A.S. Nagar, Mohali, Punjab-160062, 

India. dntripathiniper@gmail.com 


Cyclophosphamide, an alkylating agent, disturbs the oxidant and antioxidant balance that 

is associated with several unwanted toxic effects and induction of secondary cancers. 

Astaxanthin is a powerful antioxidant and possess several beneficial effects against 

various human diseases and physiological disorders. The present study was aimed to 

investigate the effects of astaxanthin against cyclophosphamide-induced oxidative stress, 

DNA damage, cell death and induction of GST-P foci in rat liver. Further attempt has 

been made to study the influence of astaxanthin on antioxidant response element (ARE) 

and the transcription factor Nrf2 (nuclear factor E(2)-related factor 2) in the induction of 

phase-II enzymes NAD(P)H: quinine oxidoreductase-1(NQO-1) and Hemoxygenase-1 

(HO-1). Both pre- and post-treatment with astaxanthin (25mg/kg) decreased 

cyclophosphamide-induced oxidative stress and DNA damage in the liver as evident from 

the restoration in malondialdehyde and glutathione level as well as modified comet assay 

parameters. Significant decrease in the number as well as area of GST-P foci in rat 

hepatocytes was observed with astaxanthin post-treatment. Treatment with astaxanthin 

significantly decreased the expression of p53 and p38 as compared to cyclophosphamide 

treated group. It was further observed that the level of Nrf2 and phase-II enzymes, i.e. 

NQO-1 and HO-1 were increased with astaxanthin treatment. The present study confirms 

that astaxanthin is a potent antioxidant and attenuates oxidative stress, DNA damage, cell 

death as well as induction of early hepatocarcinogenesis in rat induced by 

cyclophosphamide. Our results provide the evidence that one of the mechanism of 

chemoprotection offered by astaxanthin is mediated through Nrf2-ARE pathway. 

Copyright © 2009 Elsevier B.V. All rights reserved. 


Antioxidant 

59

Toxicology. 2010 Jan 12;267(1-3):147-53. Epub 2009 Nov 10. 

Effect of astaxanthin on hepatocellular 

injury following ischemia/reperfusion. 

Curek GD, Cort A, Yucel G, Demir N, Ozturk S, Elpek GO, Savas B, Aslan M. 

Source 

Department of Biochemistry, Akdeniz University Medical School, Antalya 07070, 

Turkey. 


This study investigated the effect of astaxanthin (ASX; 3,3-dihydroxybeta, beta-carotene- 

4,4-dione), a water-dispersible synthetic carotenoid, on liver ischemia-reperfusion (IR) 

injury. Astaxanthin (5 mg/kg/day) or olive oil was administered to rats via intragastric 

intubation for 14 consecutive days before the induction of hepatic IR. On the 15th day, 

blood vessels supplying the median and left lateral hepatic lobes were occluded with an 

arterial clamp for 60 min, followed by 60 min reperfusion. At the end of the experimental 

period, blood samples were obtained from the right ventricule to determine plasma 

alanine aminotransferase (ALT) and xanthine oxidase (XO) activities and animals were 

sacrificed to obtain samples of nonischemic and postischemic liver tissue. The effects of 

ASX on IR injury were evaluated by assessing hepatic ultrastructure via transmission 

electron microscopy and by histopathological scoring. Hepatic conversion of xanthine 

dehygrogenase (XDH) to XO, total GSH and protein carbonyl levels were also measured 

as markers of oxidative stress. Expression of NOS2 was determined by 

immunohistochemistry and Western blot analysis while nitrate/nitrite levels were 

measured via spectral analysis. Total histopathological scoring of cellular damage was 

significantly decreased in hepatic IR injury following ASX treatment. Electron 

microscopy of postischemic tissue demonstrated parenchymal cell damage, swelling of 

mitochondria, disarrangement of rough endoplasmatic reticulum which was also partially 

reduced by ASX treatment. Astaxanthine treatment significantly decreased hepatic 

conversion of XDH to XO and tissue protein carbonyl levels following IR injury. The 

current results suggest that the mechanisms of action by which ASX reduces IR damage 

may include antioxidant protection against oxidative injury. 

2009 Elsevier Ireland Ltd. All rights reserved. 


Antioxidant 

60

Methods Mol Biol. 2010;594:263-73. 

Assessing the neuroprotective effect of 

antioxidant food factors by application of 

lipid-derived dopamine modification 

adducts. 

Liu X, Yamada N, Osawa T. 

Source 

Laboratory of Food and Biodynamics, Graduate School of Bioagricultural Science, 

Nagoya University, Nagoya, Japan. 


Advances in understanding the neurodegenerative pathologies are creating new 

opportunities for the development of neuroprotective therapies, such as antioxidant food 

factors, lifestyle modification and drugs. However, the biomarker by which the effect of 

the agent on neurodegeneration is determined is limited. We here address hexanoyl 

dopamine (HED), one of novel dopamine adducts derived from brain polyunsaturated 

acid, referring to its in vitro formation, potent toxicity to SH-SY5Y cells, and application 

to assess the neuroprotective effect of antioxidative food factors. Dopamine is a 

neurotransmitter, and its deficiency is a characterized feature in Parkinson's disease (PD); 

thus, HED provides a new insight into the understanding of dopamine biology and 

pathophysiology of PD and a novel biomarker for the assessment of neuroprotective 

therapies. We have established an analytical system for the detection of HED and its 

toxicity to the neuroblstoma cell line, SH-SY5Y cells. Here, we discuss the 

characteristics of the system and its applications to investigate the neuroprotective effect 

of several antioxidants that originate from food. 


Antioxidant and Neuro- protection 

61

Br J Nutr. 2008 Aug;100(2):287-96. Epub 2008 Jan 11. 

Effects of apigenin, lycopene and astaxanthin on 7 betahydroxycholesterol-induced 

apoptosis and Akt 

phosphorylation in U937 cells. 

Lordan S, O'Neill C, O'Brien NM. 

Source 

Department of Food and Nutritional Sciences, University College, Cork, Republic of 

Ireland. 


Oxysterols arise from the enzymic or non-enzymic oxidation of cholesterol and have 

been shown to be cytotoxic to certain cell lines. In particular, apoptosis induced by the 

oxysterol 7 beta-hydroxycholesterol (7 beta-OH) has been associated with the generation 

of oxidative stress, cytochrome c release and caspase activation. Due to the fundamental 

importance of apoptosis in pathological processes, the identification of substances 

capable of modulating this form of cell death is now actively researched. The objective of 

the present study was to investigate if apigenin, lycopene and astaxanthin could inhibit 7 

beta-OH-induced apoptosis in U937 cells. Pretreatment with 0.1 mum-astaxanthin 

protected against apoptosis, while lycopene did not oppose the adverse effects of 7 beta- 

OH. At low concentrations, apigenin did not protect against oxysterol-induced apoptosis; 

however, at higher concentrations it intensified cell death. Additionally, we investigated 

the effect of 7 beta-OH, apigenin and astaxanthin on the activation of the serine threonine 

kinase Akt (phosphorylated Akt:Akt ratio) to determine whether the effect on cell 

viability and growth was linked to the Akt signalling pathway. Akt activation was 

decreased in the oxysterol-treated cells compared with control cells; however, this did not 

attain significance. Interestingly, activation of Akt was significantly reduced compared 

with control cells following incubation with apigenin and astaxanthin both in the absence 

and in the presence of 7 beta-OH. Our data suggest that apigenin, lycopene and 

astaxanthin failed to protect against 7 beta-OH-induced apoptosis, and the decrease in 

cell viability and the increase in apoptotic nuclei induced by the antioxidants appear to be 

associated with down regulation of Akt activity. 


Antioxidant 

62

Toxicology. 2008 Jun 27;248(2-3):96-103. Epub 2008 Mar 27. 

Astaxanthin inhibits cytotoxic and 

genotoxic effects of cyclophosphamide in 

mice germ cells. 


Source 


Education and Research, Sector-67, SAS. Nagar, Punjab 160062, India. 


Cyclophosphamide (CP), an alkylating agent used in the treatment of several cancers as 

well as an immunosuppressant in rheumatoid arthritis. It is used against several cancers 

due to its broad spectrum efficacy, but at the same time possesses unwanted risks for 

occupational exposure as well as therapy related toxicities to patients. The present study 

was aimed to investigate the protective effect of astaxanthin (AST) a red carotenoid 

pigment on CP induced germ cell toxicity in male mice. CP was administered 

intraperitoneally (i.p.) at the dose of 50, 100 and 200mg/kg body weight to mice (20-25 

g) once in a week for a period of five weeks. AST was given at the dose of 25mg/kg per 

oral (p.o.) for five consecutive days in a week for five weeks. The animals were 

sacrificed one week after the last injection of CP. The protective effect of AST against 

CP induced male germ cell toxicity was evaluated using body weight, testes and 

epididymis weight, sperm count, sperm head morphology, sperm comet assay, histology 

of testes and TUNEL assay. AST treatment significantly improved the testes weight, 

sperm count and sperm head morphology as compared to only CP treated animals. The 

result of comet assay showed that AST treatment significantly restored the sperm DNA 

damage induced by CP. Further, AST treatment showed protection against CP induced 

testicular toxicity as evident from testes histology and TUNEL assay. The present results 

indicate the chemoprotective potential of AST against CP induced germ cell toxicity in 

mice. 


Antioxidant 

63

Zhongguo Gu Shang. 2008 Mar;21(3):187-9. 

[Effects of Astaxanthin on the damage of osteoblast 

induced by H2O2]. 


Pei LP, Dong FH, Hui BD. 

Source 

Institute of Orthopaedics and Traumatology, China Academy of Chinese Medical 

Science, Beijing 100700, China. 


OBJECTIVE: To investigate the effect of Astaxanthin on enhancing the function of antioxidative 

damage in osteoblast. 

METHODS: MC3T3-E1 osteoblasts were randomly divided into five groups, including 

control group, model group, Astaxanthin group [low-dose (1 x 10(-7) mol/L), middledose 

(1 x 10(-6) mol/L), high-dose (1 x 10(-5) mol/L)], in which the activity of cells, 

activity of superoxide dismutase (SOD), the content of reactive oxygen species (ROS), 

lipid oxygen (LPO) and membrane fluidity were tested and compared. 

RESULTS: Compared with Astaxanthin groups, the activity of cells, SOD activity and 

membrane fluidity in the model group were significantly decreased (P < 0.01). However, 

the contents of ROS and LPO were significantly raised (P < 0.01). 

CONCLUSION: H2O2 can cause oxidative damage of MC3T3-E1 osteoblasts, but 

Astaxanthin can prevent or decrease its influence. 


Antioxidant 

Life Sci. 2006 Jun 6;79(2):162-74. Epub 2006 Feb 8. 

64

The effects of oral Cardax (disodium disuccinate astaxanthin) on 

multiple independent oxidative stress markers in a mouse peritoneal 

inflammation model: influence on 5-lipoxygenase in vitro and in vivo. 

Lockwood SF, Penn MS, Hazen SL, Bikádi Z, Zsila F. 

Source 

Hawaii Biotech, Inc., 99-193 Aiea Heights Drive, Suite 200, Aiea, Hawaii 96701, USA. 

slockwood@hibiotech.com 


Disodium disuccinate astaxanthin ('rac'-dAST; Cardax) is a water-dispersible C40 

carotenoid derivative under development for oral and parenteral administration for 

cardioprotection of the at-risk ischemic cardiovascular patient. In experimental infarction 

models in animals (rats, rabbits, and dogs), significant myocardial salvage has been 

obtained, up to 100% at the appropriate dose in dogs. The documented mechanism of 

action in vitro includes direct scavenging of biologically produced superoxide anion; in 

vivo in rabbits, modulation of the complement activity of serum has also been shown. A 

direct correlation between administration of the test compound in animals and reductions 

of multiple, independent markers of oxidative stress in serum was recently obtained in a 

rat experimental infarction model. For the current study, it was hypothesized that oral 

Cardax administration would inhibit oxidative damage of multiple relevant biological 

targets in a representative, well-characterized murine peritoneal inflammation model. A 

previously developed mass spectrometry-based (LC/ESI/MS/MS) approach was used to 

interrogate multiple distinct pathways of oxidation in a black mouse (C57/BL6) model 

system. In vivo markers of oxidant stress from peritoneal lavage samples (supernatants) 

were evaluated in mice on day eight (8) after treatment with either Cardax or vehicle 

(lipophilic emulsion without drug) orally by gavage at 500 mg/kg once per day for seven 

(7) days at five (5) time points: (1) baseline prior to treatment (t=0); (2) 16 h following 

intraperitoneal (i.p.) injection with thioglycollate to elicit a neutrophilic infiltrate; (3) 4 h 

following i.p. injection of yeast cell wall (zymosan; t=16 h/4 h thioglycollate+zymosan); 

(4) 72 h following i.p. injection with thioglycollate to elicit monocyte/macrophage 

infiltration; and (5) 72 h/4 h thioglycollate+zymosan. A statistically significant sparing 

effect on the arachidonic acid (AA) and linoleic acid (LA) substrates was observed at 

time points two and five. When normalized to the concentration of the oxidative 

substrates, statistically significant reductions of 8-isoprostane-F(2alpha) (8-iso-F(2alpha)) 

at time point three (maximal neutrophil recruitment/activation), and 5-HETE, 5-oxo-EET, 

11-HETE, 9-HODE, and PGF(2alpha) at time point five (maximal monocyte/macrophage 

recruitment/activation) were observed. Subsequently, the direct interaction of the 

optically inactive stereoisomer of Cardax (meso-dAST) with human 5-lipoxygenase (5- 

LOX) was evaluated in vitro with circular dichroism (CD) and electronic absorption 

(UV/Vis) spectroscopy, and subsequent molecular docking calculations were made using 

mammalian 15-LOX as a surrogate (for which XRC data has been reported). The results 

suggested that the meso-compound was capable of interaction with, and binding to, the 

solvent-exposed surface of the enzyme. These preliminary studies provide the foundation 

for more detailed evaluation of the therapeutic effects of this compound on the 5-LOX 

65

enzyme, important in chronic diseases such as atherosclerosis, asthma, and prostate 

cancer in humans. 

PMID: 

16466747 

[PubMed - indexed for MEDLINE] 

Antioxidant 

66

Anti-Inflammatory 

Mol Cells. 2003 Aug 31;16(1):97-105. 

Astaxanthin inhibits nitric oxide production and inflammatory gene 

expression by suppressing I(kappa)B kinase-dependent NF-kappaB 

activation. 

Lee SJ, Bai SK, Lee KS, Namkoong S, Na HJ, Ha KS, Han JA, Yim SV, 

Chang K, Kwon YG, Lee SK, Kim YM. 

Vascular System Research Center and Department of Molecular and Cellular 

Biochemistry, Kangwon National University Biology, Chunchon 200-701, Korea. 

Astaxanthin, a carotenoid without vitamin A activity, has shown anti-oxidant and 

anti-inflammatory activities; however, its molecular action and mechanism have 

not been elucidated. We examined in vitro and in vivo regulatory function of 

astaxanthin on production of nitric oxide (NO) and prostaglandin E2 (PGE2) as 

well as expression of inducible NO synthase (iNOS), cyclooxygenase-2, tumor 

necrosis factor-alpha (TNF-alpha), and interleukin-1beta (IL-1beta). Astaxanthin 

inhibited the expression or formation production of these proinflammatory 

mediators and cytokines in both lipopolysaccharide (LPS)-stimulated RAW264.7 

cells and primary macrophages. Astaxanthin also suppressed the serum levels of 

NO, PGE2, TNF-alpha, and IL-1beta in LPS-administrated mice, and inhibited 

NF-kappaB activation as well as iNOS promoter activity in RAW264.7 cells 

stimulated with LPS. This compound directly inhibited the intracellular 

accumulation of reactive oxygen species in LPS-stimulated RAW264.7 cells as 

well as H2O2-induced NF-kappaB activation and iNOS expression. Moreover, 

astaxanthin blocked nuclear translocation of NF-kappaB p65 subunit and 

I(kappa)B(alpha) degradation, which correlated with its inhibitory effect on 

I(kappa)B kinase (IKK) activity. These results suggest that astaxanthin, probably 

due to its antioxidant activity, inhibits the production of inflammatory mediators 

by blocking NF-kappaB activation and as a consequent suppression of IKK 

activity and I(kappa)B-alpha degradation. 


 




67

J Microbiol Biotechnol. 2008 Dec;18(12):1990-6. 

Effects of astaxanthin on the production of NO and the expression of 

COX-2 and iNOS in LPS-stimulated BV2 microglial cells. 

Choi SK, Park YS, Choi DK, Chang HI. 

Department of Biotechnology, School of Life Sciences and Biotechnology, Korea 

University, Seoul 136-701, Korea. 

Astaxanthin has shown antioxidant, antitumor, and antiinflammatory activities; 

however, its molecular action and mechanism in the nervous system have yet to 

be elucidated. We examined the in vitro effects of astaxanthin on the production 

of nitric oxide (NO), as well as the expression of inducible NO synthase (iNOS) 

and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-stimulated BV2 

microglial cells. Astaxanthin inhibited the expression or formation of nitric oxide 

(NO), iNOS and COX-2 in lipopolysaccharide (LPS)-stimulated BV-2 microglial 

cells. Astaxanthin also suppressed the protein levels of iNOS and COX-2 in LPSstimulated 

BV2 microglial cells. These results suggest that astaxanthin, probably 

due to its antioxidant activity, inhibits the production of inflammatory mediators 

by blocking iNOS and COX-2 activation or by the suppression of iNOS and 

COX-2 degradation. 



68

Biol Pharm Bull. 2004 Feb;27(2):170-3. 

Evaluation of the nitric oxide radical scavenging activity of 

manganese complexes of curcumin and its derivative. 

Sumanont Y, Murakami Y, Tohda M, Vajragupta O, Matsumoto K, 

Watanabe H. 

Department of Pharmacology, Institute of Natural Medicine, Toyama Medical and 

Pharmaceutical University, 2630 Sugitani, Toyama 930-0194, Japan. 

Curcumin manganese complex (CpCpx) and diacetylcurcumin manganese 

complex (AcylCpCpx) were determined as to their effect on the nitric oxide (NO) 

radical scavenging in vitro method using a sodium nitroprusside generating NO 

system compared with their parent compound and astaxanthin, an extreme 

antioxidant. All compounds effectively reduced the generation of NO radicals in a 

dose dependent manner. They exhibited strong NO radical scavenging activity 

with low IC(50) values. The IC(50) values of curcumin, diacetylcurcumin, CpCpx 

and AcylCpCpx obtained are 20.39+/-4.10 microM, 28.76+/-1.48 microM, 

9.79+/-1.50 microM and 8.09+/-0.99 microM, respectively. CpCpx and 

AcylCpCpx show greater NO radical scavenging than their parent compounds, 

curcumin and acetylcurcumin, respectively. However, the IC(50) values of 

curcumin and related compounds were found to be less than astaxanthin, an 

extreme antioxidant, with the lower IC(50) value of 3.42+/-0.50 microM. 


 




69

Invest Ophthalmol Vis Sci. 2003 Jun;44(6):2694-701. 

Effects of astaxanthin on lipopolysaccharide-induced inflammation 

in vitro and in vivo. 

Ohgami K, Shiratori K, Kotake S, Nishida T, Mizuki N, Yazawa K, Ohno S. 

Department of Ophthalmology and Visual Sciences, Hokkaido University 

Graduate School of Medicine, Sapporo, Japan. kohgami@med.hokudai.ac.jp 

PURPOSE: Astaxanthin (AST) is a carotenoid that is found in marine animals and 

vegetables. Several previous studies have demonstrated that AST exhibits a wide 

variety of biological activities including antioxidant, antitumor, and anti- 

Helicobacter pylori effects. In this study, attention was focused on the antioxidant 

effect of AST. The object of the present study was to investigate the efficacy of 

AST in endotoxin-induced uveitis (EIU) in rats. In addition, the effect of AST on 

endotoxin-induced nitric oxide (NO), prostaglandin E2 (PGE2), and tumor 

necrosis factor (TNF)-alpha production in a mouse macrophage cell line (RAW 

264.7) was studied in vitro. METHODS: EIU was induced in male Lewis rats by a 

footpad injection of lipopolysaccharide (LPS). AST or prednisolone was 

administered intravenously at 30 minutes before, at the same time as, or at 30 

minutes after LPS treatment. The number of infiltrating cells and protein 

concentration in the aqueous humor collected at 24 hours after LPS treatment was 

determined. RAW 264.7 cells were pretreated with various concentrations of AST 

for 24 hours and subsequently stimulated with 10 microg/mL of LPS for 24 hours. 

The levels of PGE2, TNF-alpha, and NO production were determined in vivo and 

in vitro. RESULTS: AST suppressed the development of EIU in a dose-dependent 

fashion. The anti-inflammatory effect of 100 mg/kg AST was as strong as that of 

10 mg/kg prednisolone. AST also decreased production of NO, activity of 

inducible nitric oxide synthase (NOS), and production of PGE2 and TNF-alpha in 

RAW264.7 cells in vitro in a dose-dependent manner. CONCLUSIONS: This 

study suggests that AST has a dose-dependent ocular anti-inflammatory effect, by 

the suppression of NO, PGE2, and TNF-alpha production, through directly 

blocking NOS enzyme activity. 


 

 

Comparative Study 




70

Trends Biotechnol. 2003 May;21(5):210-6. 

Haematococcus astaxanthin: applications for human health and 

nutrition. 

Guerin M, Huntley ME, Olaizola M. 

Mera Pharmaceuticals Inc., 73-4460 Queen Kaahumanu Hwy, Suite 110, Kailua- 

Kona, Hawaii 96740, USA. 

The carotenoid pigment astaxanthin has important applications in the 

nutraceutical, cosmetics, food and feed industries. Haematococcus pluvialis is the 

richest source of natural astaxanthin and is now cultivated at industrial scale. 

Astaxanthin is a strong coloring agent and a potent antioxidant - its strong 

antioxidant activity points to its potential to target several health conditions. This 

article covers the antioxidant, UV-light protection, anti-inflammatory and other 

properties of astaxanthin and its possible role in many human health problems. 

The research reviewed supports the assumption that protecting body tissues from 

oxidative damage with daily ingestion of natural astaxanthin might be a practical 

and beneficial strategy in health management. 


 

Review 



71

EFFECT OF AN ASTAXANTHIN-CONTAINING PRODUCT ON 

CARPAL TUNNEL SYNDROME 

Nir, Y., Spiller, G., Multz, C. 

Health Research and Studies Center, Los Altos, CA, 

Study Report, May, 2002 

ABSTRACT 

Carpal Tunnel Syndrome (CTS) is a debilitating disease often requiring surgery. 

Because not all patients respond to surgery and current non-surgical treatments provide 

limited benefits, investigations into alternative techniques are necessary. We investigated 

the effect of an extract of Haematococcus algae grown in Hawaii, taken three times a 

day, each dose supplying 4 mg of astaxanthin, 40 ug lutein, 65 IU vitamin A as betacarotene, 

and 50 IU of vitamin E, on the symptoms of CTS in a double-blind, placebocontrolled, 

parallel design study. Twenty participants were randomized to receive either 

the extract (13 subjects) or a placebo (7 subjects) for eight weeks. Daytime pain rate and 

duration were measured at the beginning of the study, and after 4 and 8 weeks of 

treatment, with the use of questionnaires. Results showing a trend towards decreasing 

pain rate and duration in the subjects receiving the extract, but because of the small 

number of subjects the results did not reach statistical significance (P>0.05). The 

daytime pain rates (mean ± SD) at 0, 4 and 8 weeks were, respectively, 1.69±0.99, 

1.23±0.70, and 1.00±0.88 for the treatment group, and 1.67±0.47, 1.83±0.37, and 

1.50±0.50 for the control group. Similarly, the duration of daytime pain was 2.15±1.23, 

1.69±1.13, and 1.38±1.44 for the treatment group, and 2.17± 1.07, 2.67± 1.10, and 2.17± 

1.34 for the control group. The positive trend observed in this pilot study suggests that an 

astaxanthin-containing product may be effective in treating symptoms of CTS. Further 

investigations in a larger-scale study are needed. 

Supported by a grant from the Cyanotech Corporation 


72

Effect of daily use natural astaxanthin on C-reactive protein. 

Gene A. Spiller, PhD, Antonella Dewell, MS, RD, Sally Chaves, RN, Zaga 

Rakidzich 

Health Research & Studies Center, Los Altos, CA 

Study Report, January, 2006 

ABSTRACT 

Previous studies have provided data suggesting that daily use of natural 

astaxanthin can positively address inflammatory conditions such as rheumatoid 

arthritis and carpal tunnel syndrome. In this study, the effect of daily use of 

BioAstin, a microalgae extract containing natural astaxanthin, on C-reactive 

protein was evaluated. It was found that after daily use of BioAstin for eight 

weeks C-reactive protein (CRP) was significantly lowered in the treatment group 

as compared to the placebo group. This correlation of reduced CRP and use of 

BioAstin may suggest that daily use can help reduce CRP and possibly lower 

inflammation levels in the body. 



73

ASTAXANTHIN SUPPLEMENTATION 

A.C. Fry, B.K. Schilling, L.Z.F. Chiu, N. Hori, and L.W. Weiss, FACSM. 

Human Performance Laboratories, The University of Memphis, Memphis, TN, 

USA 38152 


PURPOSE: To determine the effects of astaxanthin anti-oxidant supplementation 

as a counter-measure for delayed onset muscular soreness (DOMS) in currently 

trained individuals, nine weight trained males (X+SE: age=25.1+1.6 yrs., 

hgt=1.79+0.02 m, wgt=86.8+4.4 kg) participated in this study. METHODS: All 

subjects provided muscle biopsy samples from the vastus lateralis m. prior to 

inducing DOMS in the knee extensor mm. (10 sets x 7-10 reps, 85% eccentric 1 

RM). The subjects ingested either 4 mg.d-1 of astaxanthin (Suppl; n=4) or a 

placebo (Con; n=5) for a 3 week loading phase prior to the DOMS-inducing 

protocol, and during a 12 d recovery phase. Perceptions of DOMS at 48 hrs posteccentric 

exercise were quantified by muscle soreness ratings (0-10 Likert scale). 

Muscle fiber characteristics were determined via mATPase histochemistry and 

digital imaging to determine % cross-sectional areas of the major fiber types (I, 

IIA, IIAB/B). Due to small numbers of IIB fibers in some subjects, IIAB hybrid 

fibers were included in this fiber type population. Simple regression was used to 

determine relationships between fiber characteristics and perceptions of soreness. 

RESULTS: No differences in perceptions of soreness between the Suppl or Con 

groups were observed (p>0.05), with all subjects exhibiting a mean score of >5. 

Percent fiber type areas were similar (p>0.05) for both groups (type I, 

Suppl=47.6+8.9%, Con=41.3+2.7%; type IIA, Suppl=44.3+5.6%, 

Con=53.0+2.8%; type IIAB/B, Suppl=8.2+3.6%, Con=5.7+1.6%). However, 48 

hrs after the DOMS-inducing session, perceptions of soreness for the Suppl group 

were positively related to % area type I (r=0.90), and negatively related to % area 

types IIA (r=-0.80) and IIAB/B (r=-0.99). A distinctly different correlational 

pattern was observed for the Con group (% type I area, r=-0.58; % type IIA area, 

r=0.32; % type IIAB/B area, r=0.40). CONCLUSIONS: Collectively, these 

preliminary data suggest that astaxanthin supplementation may preferentially 

attenuate perceptions of DOMS in weight trained men with a high % area for fiber 

types IIA & AB/B. 



74

Effect of Astaxanthin on Muscular Atrophy 

Department of Exercise and Health Sciences, Faculty of Education, Yamaguchi 

University. Yamaguchi 753-8513, Japan, 2'fbyo Koso Kagaku Co. Ltd. Ureveeu, Chiba, 

279-00U. Japan, Department of Exercise PhysiaWgy, School of Health and Sports 

Science, Junteodo University, Inba, Cbibe 270-1695, Japan, Laboratory of Physiology, 

Toyobashi SOZO University, Toyohaehi 440-8511, 6 Department of Chemistry, School 

of Medicine, Juotendo University Japan, 6Hiroaaki Oakum Univcl'6ity, Hircsaki, 

Aomori 036-8577, Japan 

Objective: Patients wearing casts or other devices that hinder mobility are reported to 

have muscular atrophy. It is commonly thought that the cause is from reactive oxygen 

species (ROS). The use of Vitamin E, along with other antioxidants, prevents ROS from 

causing muscular atrophy that arises from lack of movement; however there has been 

conflicting reports. In this experiment, Astaxanthin (Ax), which is considered to be a 

more effective antioxidant than Vitamin E or beta-carotene, will be administered to 

subjects as food supplement to see its effect on muscular atrophy caused by lack of 

movement. It will also be tested if the amount of Ax intake will make a difference in its 

effectiveness. Methods: 14-week old, Wister-type, male rats were used. Mice were all 

the same weight after growth for one week under controlled conditions. The rats were 

separated into three separate groups: Control group (n=7), Ax 0.04% group, and Ax 0.2% 

group. 15 days after the administration of Ax, each rat had his right leg contained with a 

cast in an extended position to decrease muscle mass in the triceps surae muscle group 

for 10 days. At the end of the experiment, the weights of the rats were measured and, 

along with the use of Nembutal (an anesthesia), euthanized. The plantaris muscle was 

extracted for analysis. 

Results and Analysis: Groups that were administered Ax had significantly less muscle 

atrophy than those in the Control group (p

in many biological organisms, but Ax is more active than other antioxidants. Based on 

this information, we believe Ax intake prevents muscular atrophy by protecting 

membranes; preventing oxidative stress which results in atrophy; preventing the 

facilitation protease and ubiquitination. The effects due to the quantity of Ax uptake were 

not clear in this study. 


76

Long term dietary antioxidant intakes attenuate sarcopenia 

Tsubasa SHIBAGUCHl, Talmo SUGIURA, Tsukasa FURUMOTO, 

Koshiro IOUEI, Yoshiharu TlDA, Hieeebl AITOA, Kaeeumaea GOTO', 

Daijiro OHMORI, Ibshitadu YOSMOK.,V 

Oxidative stress is thought to be one of significant contributing 

factors to age-related sarccpenia. We tested the hypothesis that the long 

term dietary antioxidant (astaxanthinl intakes attenuate sarcopenie. 

Wistar strain male rats, aged 45 weeks old, were given either control (Cont) 

or astaxanthin feed (0.004%, Ax) for 1 year. 'The soleus muscle weights 

and muscle weigh-to-body weight ratios in Ax group were significantly 

heavier than in Cont group, but tibialis anterior muscle mass remained 

similar between the two dietary groups. The level of ubiquitinated 

proteins was significantly lower in soleus muscles of Ax group, but not in 

tibialis anterior muscles when compared with Cont group. Tibialis anterior 

levels of cathepsin Land caepase-S were tended to be lower in Ax group 

than in Cont group, especially significant differences observed in cathepsin 

L, whereas no differences between Cont and Ax were observed in soleus 

tbcae levels. There woro no effects ofAx supplementation on calpaia 1 and 2, 

UBC3B, CulZn SOD and nitrotyrosine levels inboth soleus and tibialis 

anterior muscles. OUT data suggest that the long term dietary 

astaxanthin intakes attenuate the age related muscle atrophy, due in part, 

to reductions in oxidative stress and ubiquitination of myofibrillar protein 

in slow soleus muscles, but not in fast tibialis anterior muscles. 


77

EFFECT OF AN ASTAXANTHIN-CONTAINING PRODUCT ON 

RHEUMATOID ARTHRITIS 

Nir, Y., Spiller, G., Multz, C. 

Health Research and Studies Center, Los Altos, CA 

Study Report, May 2002 

ABSTRACT 

Rheumatoid arthritis (RA) is a chronic destructive disorder requiring aggressive 

treatment. Conventional treatments present problems in terms of safety and efficacy, and 

the alternative therapies so far investigated have not yielded consistent results. We 

investigated the effect of an extract of Haematococcus algae grown in Hawaii, taken 

three times a day, each dose supplying 4 mg of astaxanthin, 40 ug lutein, 65 IU vitamin A 

as beta-carotene, and 50 IU of vitamin E, on the symptoms of RA in a double-blind, 

placebo-controlled, parallel design study. Twenty-one subjects were randomized to 

receive either the extract (14 subjects) or a placebo (7 subjects) for eight weeks. Pain and 

satisfaction with the ability to perform daily activities were measured at the beginning of 

the study, and after 4 and 8 weeks of treatment. The results showed a significant 

difference (P

Effect of daily use of natural astaxanthin on symptoms associated with 

Tennis Elbow (lateral humeral epicondylitis) 

Gene A. Spiller, PhD, CNS, Antonella Dewell, MS, RD, Sally Chaves, RN, Zaga 

Rakidzich, 

Health Research & Studies Center, Los Altos, CA 

Study Report, January, 2006 

ABSTRACT 

Previous studies have provided data suggesting that daily use of a microalgal extract 

containing natural astaxanthin and marketed under the trade name BioAstin® can help 

alleviate pain associated with joint damage, specifically that seen in rheumatoid arthritis 

and carpal tunnel syndrome. For this study, the benefits of daily use natural astaxanthin 

provided by BioAstin® for the purpose of alleviating pain associated with Tennis Elbow 

(lateral humeral epicondylitis) was evaluated. It was found that grip strength 

measurements (GSM) for those on the active product were significantly improved by the 

end of the study. This correlation of improved GSM and use of natural astaxanthin may 

suggest that daily use can help alleviate pain associated with Tennis Elbow, and increase 

mobility. This improvement may greatly improve the standard of living for those who 

suffer from such joint disorders. 



79

Am J Cardiol 2008;101[suppl]:58D– 68D 

Astaxanthin: A Novel Potential Treatment for Oxidative Stress and 

Inflammation in Cardiovascular Disease 

Fredric J. Pashkow, MD,a,b,* David G. Watumull,b and Charles L. 

Campbell, MDc 

Oxidative stress and inflammation are implicated in several different 

manifestationsof cardiovascular disease (CVD). They are generated, in part, from 

the overproduction of reactive oxygen species (ROS) and reactive nitrogen 

species (RNS) thatactivate transcriptional messengers, such as nuclear factor–_B, 

tangibly contributing to endothelial dysfunction, the initiation and progression of 

atherosclerosis, irreversible damage after ischemic reperfusion, and even 

arrhythmia, such as atrial fibrillation. Despite this connection between oxidative 

stress and CVD, there are currently no recognized therapeutic interventions to 

address this important unmet need. Antioxidants that provide a broad, “upstream” 

approach via ROS/RNS quenching or free radical chain breaking seem an 

appropriate therapeutic option based on epidemiologic, dietary, and in vivo 

animal model data. However, human clinical trials with several different wellknown 

agents, such as vitamin E and _-carotene, have been disappointing. Does 

this mean antioxidants as a class are ineffective, or rather that the “right” 

compound(s) have yet to be found, their mechanisms of action understood, and 

their appropriate targeting and dosages determined? A large class of potent 

naturally-occurring antioxidants exploited by nature—the oxygenated carotenoids 

(xanthophylls)— have demonstrated utility in their natural form but have eluded 

development as successful targeted therapeutic agents up to the present time. This 

article characterizes the mechanism by which this novel group of antioxidants 

function and reviews their preclinical development. Results from multiple species 

support the antioxidant/anti-inflammatory properties of the prototype compound, 

astaxanthin, establishing it as an appropriate candidate for development as a 

therapeutic agent for cardiovascular oxidative stress and inflammation. 


80

Phytother Res. 2010 Jul 14. [Epub ahead of print] 

Summative interaction between astaxanthin, Ginkgo biloba extract (EGb761) and 

vitamin C in Suppression of respiratory inflammation: a comparison with 

ibuprofen. 

Haines DD, Varga B, Bak I, Juhasz B, Mahmoud FF, Kalantari H, Gesztelyi R, Lekli I, 

Czompa A, Tosaki A. 

Department of Pharmacology, Faculty of Pharmacy, University of Debrecen, Debrecen, 

Hungary. 


In this study, combinations of Ginkgo biloba leaf extract (EGb761) plus the carotenoid 

antioxidant astaxanthin (ASX) and vitamin C were evaluated for a summative dose effect 

in the inhibition of asthma-associated inflammation in asthmatic guinea-pigs. Ovalbuminsensitized 

Hartley guinea-pigs challenged with ovalbumin aerosol to induce asthma, were 

administered EGb761, ASX, vitamin C or ibuprofen. Following killing, bronchoalveolar 

lavage (BAL) fluid was evaluated for inflammatory cell infiltrates and lung tissue cyclic 

nucleotide content. Each parameter measured was significantly altered to a greater degree 

by drug combinations, than by each component acting independently. An optimal 

combination was identified that included astaxanthin (10 mg/kg), vitamin C (200 mg/kg) 

and EGb761 (10 mg/kg), resulting in counts of eosinophils and neutrophils each 1.6-fold 

lower; macrophages 1.8-fold lower, cAMP 1.4-fold higher; and cGMP 2.04-fold higher 

than levels in untreated, asthmatic animals (p < 0.05). In conclusion, EGb761, ASX and 

vitamin C are shown here to interact summatively to suppress inflammation with efficacy 

equal to or better than ibuprofen, a widely used non-steroidal antiinflammatory drug 

(NSAID). Such combinations of non-toxic phytochemicals constitute powerful tools for 

the prevention of onset of acute and chronic inflammatory disease if consumed regularly 

by healthy individuals; and may also augment the effectiveness of therapy for those with 

established illness. Copyright (c) 2010 John Wiley & Sons, Ltd. 



81

J Microbiol Biotechnol. 2008 Dec;18(12):1990-6. 

Effects of astaxanthin on the production 

of NO and the expression of COX-2 and 

iNOS in LPS-stimulated BV2 microglial 

cells. 

Choi SK, Park YS, Choi DK, Chang HI. 

Source 


University, Seoul 136-701, Korea. 


Astaxanthin has shown antioxidant, antitumor, and antiinflammatory activities; however, 

its molecular action and mechanism in the nervous system have yet to be elucidated. We 

examined the in vitro effects of astaxanthin on the production of nitric oxide (NO), as 

well as the expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) 

in lipopolysaccharide (LPS)-stimulated BV2 microglial cells. Astaxanthin inhibited the 

expression or formation of nitric oxide (NO), iNOS and COX-2 in lipopolysaccharide 

(LPS)-stimulated BV-2 microglial cells. Astaxanthin also suppressed the protein levels of 

iNOS and COX-2 in LPS-stimulated BV2 microglial cells. These results suggest that 

astaxanthin, probably due to its antioxidant activity, inhibits the production of 

inflammatory mediators by blocking iNOS and COX-2 activation or by the suppression 

of iNOS and COX-2 degradation. 



82

J Biol Chem. 2009 Oct 9;284(41):28172-9. Epub 2009 Aug 21. 

Inhibitory effect of carotenoids on the 

degranulation of mast cells via 

suppression of antigen-induced 

aggregation of high affinity IgE receptors. 

Sakai S, Sugawara T, Matsubara K, Hirata T. 

Source 

Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, 

Kyoto 606-8502, Japan. 


Carotenoids have been demonstrated to possess antioxidative and anti-inflammatory 

effects. However, there is no report that the effects of carotenoids on degranulation of 

mast cell is critical for type I allergy. In this study, we focused on the effect of 

carotenoids on antigen-induced degranulation of mast cells. Fucoxanthin, astaxanthin, 

zeaxanthin, and beta-carotene significantly inhibited the antigen-induced release of betahexosaminidase 

in rat basophilic leukemia 2H3 cells and mouse bone marrow-derived 

mast cells. Those carotenoids also inhibited antigen-induced aggregation of the high 

affinity IgE receptor (Fc epsilonRI), which is the most upstream of the degranulating 

signals of mast cells. Furthermore, carotenoids inhibited Fc epsilonRI-mediated 

intracellular signaling, such as phosphorylation of Lyn kinase and Fyn kinase. It suggests 

that the inhibitory effect of carotenoids on the degranulation of mast cells were mainly 

due to suppressing the aggregation of Fc epsilonRI followed by intracellular signaling. In 

addition, those carotenoids inhibited antigen-induced translocation of Fc epsilonRI to 

lipid rafts, which are known as platforms of the aggregation of Fc epsilonRI. We assume 

that carotenoids may modulate the function of lipid rafts and inhibit the translocation of 

Fc epsilonRI to lipid rafts. This is the first report that focused on the aggregation of Fc 

epsilonRI to investigate the mechanism of the inhibitory effects on the degranulation of 

mast cells and evaluated the functional activity of carotenoids associated with lipid rafts. 



83

Eur J Nutr. 2010 Mar;49(2):119-26. Epub 2009 Sep 26. 

Astaxanthin suppresses scavenger receptor expression and 

matrix metalloproteinase activity in macrophages. 

Kishimoto Y, Tani M, Uto-Kondo H, Iizuka M, Saita E, Sone H, Kurata H, Kondo K. 

Source 

Institute of Environmental Science for Human Life, Ochanomizu University, Tokyo, 

Japan. 


BACKGROUND: Astaxanthin is a red carotenoid pigment which has significant 

potential for antioxidant activity. The macrophages in atherosclerotic lesions, known as 

activated macrophages, express scavenger receptors responsible for the clearance of 

pathogenic lipoproteins. In addition, the expression and secretion of proteolytic enzymes, 

matrix metalloproteinases (MMPs), and pro-inflammatory cytokines are remarkably 

promoted in activated macrophages. 

AIM OF THE STUDY: In this study, we investigated the effects of astaxanthin on 

the expression of scavenger receptors, MMPs, and pro-inflammatory cytokines in 

macrophages. 

METHODS: THP-1 macrophages were incubated with 5-10 microM astaxanthin for 

24 h. The expression levels of scavenger receptors, MMPs, and pro-inflammatory 

cytokines were determined by Western blot analysis or real-time RT-PCR. The MMP-9 

and -2 activities were examined by gelatin zymography and total MMP activity was 

measured by fluorometry. 

RESULTS: We found that astaxanthin remarkably decreased the class A scavenger 

receptor and CD36 expression in the protein and mRNA levels. Astaxanthin also reduced 

MMP-1, -2, -3, -9, -12, and -14 activity and expression. The mRNA expression of tumor 

necrosis factor-alpha, interleukin-1beta, interleukin-6, inducible nitric oxide synthase, 

and cyclooxygenase-2 were significantly suppressed by astaxanthin. Furthermore, 

astaxanthin inhibited the phosphorylation of nuclear factor-kappaB. 

CONCLUSIONS: These results indicate that astaxanthin has inhibitory effects on 

macrophage activation, such as scavenger receptors up-regulation, MMPs activation, and 

pro-inflammatory cytokines secretion. 



84

Skin Health 

Carotenoid Science Vol 10, p 91-5 (2006) 

The Effects of a Dietary Supplement Containing Astaxanthin on Skin 

Condition 

Eiji Yamashita 

The somatic effect son human skin by 4mg per day astaxanthin supplementation were 

demonstrated in a single blind placebo controlled study using forty-nine US healthy 

middle-aged woman. There were significant improvements in fine lines/wrinkles and 

elasticity by dermatologist’s assessment and in the moisture content by instrumental 

assessment at week 6 compares to base-line initial values. 

Astaxanthin, widely and naturally distributed in marine organisms, including Crustacea 

such as shrimps and crabs and such fish as salmon and sea bream exhibits a strong antioxidative 

effect, and its action is reported to 1,000 times stronger then alpha-tocopherol 

and approximately 40 times stronger than beta-carotene. It has also been reported that 

astaxanthin doesn’t have any pro-oxidative nature like beta-carotene and lycopene and its 

potent anti-oxidant property is exhibited at the cell membrane. Although used only as a 

coloring in the past (either as a food additive or a dye-up agent for cultured fish), 

astaxanthin has become one of the major materials eagerly anticipated by industries for 

dietary supplements and personal care products. 

Furthermore its other various important benefits to date have suggested fro human 

health such as anti-inflammation, LDL cholesterol oxidation suppression, 

immunomodulation, anti-stress, limiting diabetic nephropathy, improved semen quality, 

attenuating eye fatigue, sport performance and endurance, limiting exercised induced 

muscle damage and improving hypertension. 

In terms of dermatolofical actions, suppression of hyper-pigmentation, inhibitions 

of melanin synthesis and photo-aging have been reported. We have also reported visual 

wrinkled reduction by topical astaxanthin. However, only one study for internal use 

about cosmetic benefit of a dietary supplement including astaxanthin and tocotrienol on 

human skin has been reported. 

Here we report the effects of a dietary supplement containing astaxanthin on skin 

condition performed in the United States of America. 

Skin Health 

85

Exp Dermatol. 2009 Mar;18(3):222-31. Epub 2008 Sep 18. 

Astaxanthin, canthaxanthin and beta-carotene differently affect 

UVA-induced oxidative damage and expression of oxidative stressresponsive 

enzymes. 

Camera E, Mastrofrancesco A, Fabbri C, Daubrawa F, Picardo M, Sies H, 

Stahl W. 

Laboratorio di Fisiopatologia Cutanea, Istituto Dermatologico San Gallicano 

(IRCCS), Rome, Italy. camera@ifo.it 

Carotenoids are used for systemic photoprotection in humans. Regarding 

mechanisms underlying photoprotective effects of carotenoids, here we compared 

the modulation of UVA-related injury by carotenoids. Human dermal fibroblasts 

(HDF) were exposed to moderate doses of UVA, which stimulated apoptosis, 

increased levels of reactive oxygen species and thiobarbituric acid reactive 

substances, decreased antioxidant enzymes activities, promoted membrane 

perturbation, and induced the expression of heme oxygenase-1 (HO-1). The 

carotenoids astaxanthin (AX), canthaxanthin (CX) and beta-carotene (betaC) were 

delivered to HDF 24 h before exposure to UVA. Astaxanthin exhibited a 

pronounced photoprotective effect and counteracted all of the above-mentioned 

UVA-induced alterations to a significant extent. beta-Carotene only partially 

prevented the UVA-induced decline of catalase and superoxide dismutase 

activities, but it increased membrane damage and stimulated HO-1 expression. 

Moreover, betaC dose-dependently induced caspase-3 activity following UVA 

exposure. In contrast, CX had no effect on oxidative damage, except for HO-1 

expression, which was augmented. Uptake of AX by fibroblasts was higher than 

that of the other two carotenoids. The photostability of the three compounds in 

fibroblasts was AX > CX >> betaC. The data indicate that the oxo-carotenoid AX 

has a superior preventive effect towards photo-oxidative changes in cell culture. 



Skin Health 

86

J Dermatol Sci. 2002 Oct;30(1):73-84. 

Modulatory effects of an algal extract containing astaxanthin on 

UVA-irradiated cells in culture. 

Lyons NM, O'Brien NM. 

Department of Food Science, Food Technology and Nutrition, University College 

Cork, Cork, Ireland. nob@ucc.ie 

UV radiation from sunlight is the most potent environmental risk factor in skin 

cancer pathogenesis. In the present study the ability of an algal extract to protect 

against UVA-induced DNA alterations was examined in human skin fibroblasts 

(1BR-3), human melanocytes (HEMAc) and human intestinal CaCo-2 cells. The 

protective effects of the proprietary algal extract, which contained a high level of 

the carotenoid astaxanthin, were compared with synthetic astaxanthin. DNA 

damage was assessed using the single cell gel electrophoresis or comet assay. In 

1BR-3 cells, synthetic astaxanthin prevented UVA-induced DNA damage at all 

concentrations (10 nM, 100 nM, 10 microM) tested. In addition, the synthetic 

carotenoid also prevented DNA damage in both the HEMAc and CaCo-2 cells. 

The algal extract displayed protection against UVA-induced DNA damage when 

the equivalent of 10 microM astaxanthin was added to all three-cell types, 

however, at the lower concentrations (10 and 100 nM) no significant protection 

was evident. There was a 4.6-fold increase in astaxanthin content of CaCo-2 cells 

exposed to the synthetic compound and a 2.5-fold increase in cells exposed to 

algal extract. In 1BR-3 cells, exposure to UVA for 2 h resulted in a significant 

induction of cellular superoxide dismutase (SOD) activity, coupled with a marked 

decrease in cellular glutathione (GSH) content. However pre-incubation (18 h) 

with 10 microM of the either the synthetic astaxanthin or the algal extract 

prevented UVA-induced alterations in SOD activity and GSH content. Similarly, 

in CaCo-2 cells a significant depletion of GSH was observed following UVAirradiation 

which was prevented by simultaneously incubating with 10 microM of 

either synthetic astaxanthin or the algal extract. SOD activity was unchanged 

following UVA exposure in the intestinal cell line. This work suggests a role for 

the algal extract as a potentially beneficial antioxidant. 



Skin Health 

87

J Dermatol Sci. 1998 Mar;16(3):226-30. 

Modulation of UVA light-induced oxidative stress by beta-carotene, 

lutein and astaxanthin in cultured fibroblasts. 

O'Connor I, O'Brien N. 

Department of Nutrition, University College, Cork, Ireland. 

The ability of beta-carotene, lutein or astaxanthin to protect against UVA-induced 

oxidative stress in rat kidney fibroblasts (NRK) was assessed. Activities of the 

antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD), and 

changes in thiobarbituric acid reactive substances (TBARS) were measured as 

indices of oxidative stress. Exposure to UVA light at a dose intensity of 5.6 

mW/cm2 for 4 h resulted in a significant decrease in CAT and SOD activities and 

a significant increase in TBARS. No cytotoxicity, as indicated by lactate 

dehydrogenase (LDH) release, was observed. beta-Carotene (1 microM), lutein (1 

microM) and astaxanthin (10 nM) protect against UVA light-induced oxidative 

stress in vitro with astaxanthin exhibiting superior protective properties. 



Skin Health 

88

Int J Vitam Nutr Res. 1995;65(2):79-86. 

Vitamin A status and metabolism of cutaneous polyamines in the 

hairless mouse after UV irradiation: action of beta-carotene and 

astaxanthin. 

Savouré N, Briand G, Amory-Touz MC, Combre A, Maudet M, Nicol M. 

Biochimie Médicale A - Faculté de Médecine de Rennes, France. 

Solar radiations (UV A and B) can cause epidermis photoaging and skin cancers. 

These frequently irreversible effects result from the in situ generation of free 

radicals. However, it has been noted that nutritional factors can modulate 

photochemical damage, in particular the common carotenoids present in food, 

which can be considered as potential prophylactic agents against carcinogenesis. 

We investigated the effect of UV A and B radiations on the skin of the SKH1 

hairless mouse fed a diet either lacking in vitamin A or supplemented with retinol, 

beta-carotene or astaxanthin. The latter is an oxygenated carotenoid (like 

canthaxanthin) without provitamin A activity and with strong singlet oxygen 

quenching ability. After analysing of vitamin status of each group (plasma retinol 

concentrations and hepatic reserves), we searched for UV-induced modifications 

of polyamine metabolism by measuring epidermal ornithine decarboxylase (ODC) 

activity and free polyamines concentration (putrescine, spermidine and spermine). 

In the basal state without irradiation, differences in ODC activity between groups 

were nonsignificant; but after UV stimulation, ODC increased markedly in the 

skin of vitamin A-deficient animals, much more than in other groups. Curiously, 

the addition of astaxanthin or beta-carotene to the regimen containing retinol 

reduced the protective effect of retinol alone. Regarding polyamines after 

irradiation, putrescine was significantly increased in the skin of deficient animals, 

in parallel with ODC activity. However, astaxanthin had a stronger inhibitory 

effect on putrescine accumulation than retinol, and decreased spermidine and 

spermine concentrations: this suggests a specific action on transglutaminases. 



Skin Health 

89

Beauty From Within: A Synergistic Combination Of Astaxanthin And 

Tocotrienol For Beauty Supplements 

Yamashita, E. 

(2002) Cosmetic Benefit of Dietary Supplements Containing Astaxanthin and 

Tocotrienol on Human Skin. Food Style 21 6(6):112-17. 

Previously reported dermatological benefits of natural astaxanthin included antihyperpigmentation, 

melanin synthesis inhibition, and reduced photo-skin aging. 

Hence, the potency of astaxanthin for cosmetic effect is “clearly visible”. 

Another class of natural compounds called tocotrienols also offer cosmetic 

benefits. A member of the vitamin E family, its isomeric form (chemically 

identical, but structurally different) imparts greater protection against free radicals 

than its popular cousin, alpha-tocopherol. Tocotrienols are generally 40-60 times 

more powerful than alpha-tocopherols in terms of free radical protection. Both 

astaxanthin and tocotrienols are found naturally in daily foods we consume. By 

concentrating these into an oral beauty supplement, it can provide an excellent 

source of protection in addition to the daily skincare regime. Results in 4 weeks 

supplementation indicated reduction in fine wrinkles, increased skin moisture and 

increased skin elasticity compared to placebo. 

Skin Health 

90

(Adapted from Nutrition Business Journal, December 2004) 

Beauty clinical: Astaxanthin with Omega 3 and Marine 

Glycosaminoglycans 

Alain Thibodeau, Director of Scientific Affairs for Atrium Biotechnologies Inc. in 

Quebec, Canada published results of a blinded parallel group clinical trial on 

topical and supplemental forms of a product they call MRT2 (Matrix 

Rejuvenation Technology 2). The trial was done using both a topical product 

containing marine glycosaminoglycans and a supplement containing marine 

glycosaminoglycans, astaxanthin and omega-3 fatty acids. The trial involved 100 

subjects. 

Significant improvements were measured in skin hydration and elasticity. Skin 

appearance (including skin tone, fine lines and sallowness) also showed benefits, 

with the strongest improvements made in subjects using both the supplement and 

the topical products. 

“We can demonstrate a synergistic activity between the topical product and the 

dietary supplement…The topical product works. The supplement works as well, 

but you get much better results from using both” said Thibodeau. 

Skin Health 

91

Cosmetics.! Toiletries"'magazine/57 VOL lIS, No. l/January2003 57-9 

Dietary /Nutritional Supplements: The New Allyto TopicalCosmetic 

Formulations? 

Alain Thibodeau and Edouard lauzler 

Dietary/Nutritional Supplements 

Dietary/nutritional supplements can be used to make active nutrients available to 

all organs of the body. As mentioned earlier, skin is an organ and may therefore 

benefit from active nutrients conveyed by dietary/nutritional supplements. The 

repercussions of nutrient on skin health are well exemplified by the fact that some 

skin disorders are directly linked to nutritional deficiencies. 

Conversely, skin plays a major role in maintaining bone health through the 

synthesis of vitamin D. the interrelation between skin and the nutritional 

homeostasis has been recently highlighted and calls upon the understanding of the 

cellular and molecular processes in play. 

We have performed a clinical trail in which a topical cream formulation 

and a dietary/nutritional supplement were concomitantly administered. The 

dietary/nutritional supplement provided proteoglycans, collagen, glucosamine, 

carotenoid pigment (astaxanthin esters) and omega-3 essential fatty acids (EPA 

and DHA). The efficacy of this regimen was demonstrated on the visual 

appearance of signs of aging as well as by the amelioration of functional 

properties of the skin. 

Skin Health 

92

Journal of Cosmetic Dermatology Volume 4 Page 277 - December 2005 

A novel micronutrient supplement in skin aging: a randomized 

placebo-controlled double-blind study 

Alain Béguin 

Summary 

Background: Skin aging, a combination of intrinsic and environmentally induced 

processes, predominantly ultraviolet (UV) light from the sun, results in 

characteristic tissue alterations, such as the degradation of collagen and the 

formation of visible fine lines and wrinkles. 

Objective To test the efficacy and safety of a novel micronutrient supplement 

(Estime® containing BioAstin Natural Astaxanthin) in skin aging. 

Methods A 4-month randomized double-blind controlled study including 40 

subjects where the supplement was tested against placebo for 3 months followed 

by a 1-month supplement-free period for both groups to assess lasting effects. 

Efficacy measurements included skin surface evaluation, ultrasound measurement 

of sun-exposed and protected areas of the skin (back of the hand and ventral 

forearms, respectively), and photographic assessment. 

Results All investigated parameters showed a continuous and significant 

improvement in the active group during the 3 months of supplementation as 

compared to placebo. Photographs showed visible improvement of the overall 

skin appearance and reduction of fine lines. Ultrasound measurements showed an 

increase in dermis density of up to 78% in the active group (P < 0.0001). The 

final assessment after 1 month without supplementation showed no further 

improvements, but a slight decrease was observed in most improved parameters. 

No treatment-related side effects were reported. 

Conclusion The study demonstrated that the supplement appears to be effective 

and safe as an oral supplement to protect the skin and support its repair process. 

Recommendations are made for further evaluations. 

Skin Health 

93

Fragr J VOL.34;NO.3;PAGE.21-27(2006) 

Biological activities of astaxanthin and its cosmeceutical application. 

YAMASHITA EIJI 

The present review covers cosmeceutical benefits of astaxanthin that is one of the 

most abundant carotenoids in nature, particularly in marine based life. The antioxidant 

properties of astaxanthin without any pro-oxidative nature working at cell 

membrane and cosmeceutical effects such as anti-hyperpigmentation, antiphotoaging, 

melanin inhibition and visual wrinkle reduction by topical or internal 

use and one of the action mechanisms of astaxanthin on NF-kB dependent 

inflammation are introduced. And current and future cosmeceutical applications 

of astaxanthin particularly from a green microalgae Haematococcus pluvialis that 

is the most ideal source in the earth are discussed describing actual examples of 

astaxanthin containing skin care products in Japanese market. 

Skin Health 

94

Photoprotective Effect of Astaxanthin Applied to the Skin 

Arakane, K. 2002. KOSE Corporation 

Reactive oxygen species generated by exposing the skin to sunlight are 

responsible for sunburn, lipid peroxidation and degenerative changes in dermal 

connective tissues. This causes premature aging of the skin. 

A researcher from a Japanese company called KOSE Corporation compared 

astaxanthin to other commonly used ingredients in cosmetics that are thought to 

protect the skin from the damaging effects of sunlight. He found that astaxanthin 

potentially offers greater antioxidant protection against premature signs of aging. 

Skin Health 

95

Carotenoid Science, Vol. 5, p21-4 April 2002 Toyama, Japan 

Superior Skin Protection via Astaxanthin 

Kumi Arakane 

It has been believed for a long time that the skin exists only for the purpose of 

merely protecting our body by physically shielding it from outside factors. But in 

recent years, along with the radical progress in the field of dermatological science 

studies, it is known that the skin does actually indicate various responses and 

accept acute and chronic damages under UV irradiation. According to the 

enthusiastic studies to clarify the mechanism leading to the skin damages, 

nowadays the reactive oxygen species generated by UV irradiation is considered 

to be an important factor mediating photo-induced skin damages. Accumulation 

skin damages by reactive oxygen species such' as lipid peroxidation, sunburn and 

degenerative changes in dermal connective tissues induce the skin aging. To 

protect skin from reactive oxygen species, many cosmetics contain nowadays 

both naturally occurring molecules and synthetic compounds as antioxidant 

However. Β-carotene was the only carotenoid for cosmetics among more than 600 

carotenoids which had been isolated from nature, until astaxanthin from Antarctic 

krill was approved for cosmetics in 1997. In this paper, I would like to show the 

possibility of astaxanthin as a cosmetic ingredient and the useful formula for 

maintaining the stability of astaxanthin in the preparation. 

Skin Health 

96

Journal of Japanese Cosmetic Science Society VOL.29;NO.1;PAGE.9-19(2005) 

Preventive Effects of Carotenoids on Photoaging and Its Application 

for Cosmetics 

MIZUTANI YUKI; SAKATA OSAMU; HOSHINO TAKU; HONDA 

YOSHIKO; YAMASHITA MIKA; ARAKANE KUMI; SUZUKI TADASHI 

Carotenoids are functional materials and more than 650 kinds of carotenoides are 

isolated from nature. They have been applied for foods, but most of these 

carotenoids have not been studied in terms of their effects on skin functions, and 

because of their instability under light exposure they were hardly used in the 

cosmetics field until now. Using hairless mice irradiated with UVB to produce 

photoaged skin, we investigated the inhibitory effect of astaxanthin on wrinkle 

formation, decrease of skin elasticity, ultrastructural change of dermal collagen 

fiber bundles and elastic fibers and the level of matrix metalloproteinase-1 

(MMP-1) activity. These results indicated that the astaxanthin had the superior 

protection effect on photoaging as a ROS scavenger. It is well known that 

carotenoids are easy to decompose during storage by UV light and oxygen. We 

found that the incorporation of dl-.ALPHA.-tocopherol and .ALPHA.-glucosyl 

rutin was able to maintain long-term stability of astaxanthin in preparation. This 

research demonstrated the superior anti-aging effects by carotenoids and this is 

the first time for carotenoids to be practically applicable to cosmetic formulation. 

Skin Health 

97

Fragr J VOL.29;NO.12;PAGE.98-103(2001) 

Effects of astaxanthin from Haematococcus pluvialis on human skin. 

Patch testing Skin repeated application test Effect on wrinkle 

reduction. 

SEKI TAISUKE; SUEKI HIROHIKO; KONO HIROMI; SUGANUMA 

KAORU; YAMASHITA EIJI 

Astaxanthin is a natural color carotenoid found in salmon, salmon eggs, krill, and 

crab. Therefore, astaxanthin has been contained in the human diet for a long time. 

Astaxanthin from krill has been used for cosmetics to suppress post-UVB 

hyperpigmentation in human skin and food color additives. Recently, astaxanthin 

from Haematococcus pluvialis is available using new fermentation technology of 

H. pluvialis and it is used for dietary supplements, food color additives and 

cosmetics. Effects of astaxanthin from Haematococcus pluvialis on human 

subjects were tested. No serious adverse effects were observed by patch testing 

and sequencing applied test on human skin. In a pilot study, the skin repeated 

application test of cream containing astaxanthin on human skin showed the visual 

wrinkle reduction. The present paper described about patch testing, skin repeated 

application test, and a pilot study evaluating the wrinkle reduction effect on 

human skin. 

Skin Health 

98

Journal of Japanese Cosmetic Science Society VOL.27;NO.4;PAGE.298-303(2003) 

Effect of Antioxidant to Inhibit UV-Induced Wrinkles 

ARAKANE KUMI 

Living organisms are protected from harmful ultraviolet (UV) rays by the ozone 

layer surrounding the earth. However, depletion of the ozone layer and an 

increase in the amount of UV rays in sunlight reaching the earth's surface have 

been recently reported. As a result, social concerns over the effects of UV on 

living organisms have been increasing year by year. The skin covers the outer 

surface of the body, and so it is most vulnerable to UV. Because UV-induced 

wrinkles are prominently observed only in sun-exposed areas, they are apparently 

caused by chronic damage due to accumulated UV exposure. In addition to a 

change in appearance (large deep wrinkles), histological changes including 

thickening of the epidermis and dermis, elastin fiber deposition and decreased 

collagen fibers are observed as a result of continuous UV irradiation. Many 

reports indicate the involvement of action of reactive oxygen species in UVinduced 

wrinkles formation. Reactive oxygen species are known to damage 

essential elements including collagen and elastin which maintain elasticity and 

firmness of the skin, and also damage the function of fibroblasts producing these 

elements. It goes without saying that application of UV-absorbing agents is 

effective in preventing changes associated with photoaging. It is also reported that 

antioxidants such as vitamins C, E and iron chelators are effective for photoaging. 

We demonstrate that reactive oxygen species quenchers play an important role in 

reduction of UV-induced wrinkles formation using a carotenoid, astaxanthin, 

which has no pro-vitamin A activity unlike .BETA.-carotene, and a new iron 

chelator, N-(4-pyridoxylmethylene)-L-serine (PYSer), which consists of 

biomimetic molecules and effectively suppresses production of hydroxyl radical 

by chelating iron in skin. The demonstrable and potential roles of antioxidants for 

suppression of UV-induced wrinkles formation effectively are summarized here. 

Skin Health 

99

J Dermatol Sci. 2010 May;58(2):136-42. Epub 2010 Feb 18. 

Astaxanthin attenuates the UVA-induced up-regulation of matrixmetalloproteinase-1 

and skin fibroblast elastase in human dermal 

fibroblasts. 

Suganuma K, Nakajima H, Ohtsuki M, Imokawa G. 

Department of Dermatology, Jichi Medical University, 3311-1 Yakushiji, Shimotuke, 

Tochigi 329-0498, Japan. 


BACKGROUND: Repetitive exposure of the skin to UVA radiation elicits sagging more 

frequently than wrinkling, which is mainly attributed to its biochemical mechanism to upregulate 

the expression of matrix-metalloproteinase (MMP)-1 and skin fibroblast elastase 

(SFE)/neutral endopeptidase (NEP), respectively. OBJECTIVE: In this study, we 

examined the effects of a potent antioxidant, astaxanthin (AX), on the induction of MMP- 

1 and SFE by UVA treatment of cultured human dermal fibroblasts. METHODS: Those 

effects were assessed by real-time RT-PCR, Western blotting and enzymic activity 

assays. RESULTS: UVA radiation elicited a significant increase in the gene expression 

of MMP-1 as well as SFE/NEP (to a lesser extent) which was followed by distinct 

increases in their protein and enzymatic activity levels. The addition of AX at 

concentrations of 4-8 microM immediately after UVA exposure significantly attenuated 

the induction of MMP-1 and SFE/NEP expression elicited by UVA at the gene, protein 

and activity levels although both the UVA stimulation and the subsequent AX inhibition 

were greater for MMP-1 than for SFE/NEP. Analysis of the UVA-induced release of 

cytokines revealed that UVA significantly stimulated only the secretion of IL-6 among 

the cytokines tested and that AX significantly diminished only the IL-6 secretion. 

CONCLUSION: These findings indicate that, based on different effective concentrations 

of AX, a major mode of action leading to the inhibition elicited by AX depends on 

inhibition of UVA effects of the reactive oxygen species-directed signaling cascade, but 

not on interruption of the IL-6-mediated signaling cascade. We hypothesize that AX 

would have a significant benefit on protecting against UVA-induced skin photo-aging 

such as sagging and wrinkles. 2010 Japanese Society for Investigative Dermatology. 

Published by Elsevier Ireland Ltd. All rights reserved. 


Skin Health 

100

Plant Physiol Biochem. 2008 Oct;46(10):875-83. Epub 2008 Jun 6. 

Transgenic carrot plants accumulating ketocarotenoids show 

tolerance to UV and oxidative stresses. 

Jayaraj J, Punja ZK. 

Department of Biological Sciences, Simon Fraser University, 8888 University 

Drive, Burnaby, BC , Canada. jaya@sfu.ca 

Ketocarotenoids are strong antioxidant compounds which accumulate in salmon, 

shrimp, crustaceans and algae, but are rarely found naturally in higher plants. In 

this study, we engineered constitutive expression of an algal beta-carotene 

ketolase gene (bkt) in carrot plants to produce a number of ketocarotenoids from 

beta-carotene. These included astaxanthin, adonirubin, canthaxanthin, 

echinenone, adonixanthin and beta-cryptoxanthin. Leaves accumulated up to 56 

microg/g total ketocarotenoids and contained higher beta-carotene levels but 

lower levels of alpha-carotene and lutein. The photosynthetic capacity of 

transgenic plants was not significantly altered by these changes. However, when 

high-expressing transgenic plants were exposed to UV-B irradiation, they grew 

significantly better than the wild-type controls. Similarly, leaf tissues exposed to 

various oxidative stresses including treatment with H(2)O(2) and methyl viologen 

showed less injury and retained higher levels of chlorophyll a+b. Total carotenoid 

extracts from transgenic leaves had higher antioxidant and free-radical scavenging 

activity in vitro compared to control leaves. Transgenic tissues also accumulated 

lower amounts of H(2)O(2) following exposure to oxidative stresses, suggesting 

that free radical and reactive oxygen species were quenched by the 

ketocarotenoids. 



Skin Health 

101

Eye Health 

Japanese Journal of Clinical Ophthalmology VOL.58;NO.6;PAGE.1051-1054(2004) 

Changes in visual function following peroral astaxanthin 

NAKAMURA AKIRA; ISOBE RYOKO; OTAKA YASUHIRO; ABEMATSU 

YASUKO; NAKATA DAISUKE; HONMA CHIKA ; SAKURAI SHIZUKA; 

SHIMADA YOSHIAKI; HORIGUCHI MASAYUKI 

We evaluated the effect of astaxanthin on visual function in 49 eyes of 49 healthy 

volunteers. They were over 40 years of age. They were divided into 4 groups 

matched for age and gender. Each group was given peroral astaxanthin once a 

day. The dosage was 0mg, 2mg, 4mg, or 12mg for each group. After ingestion of 

astaxanthin for consecutive 28 days, the uncorrected far visual acuity significantly 

improved in groups receiving 4mg or 12mg. The accommodation time 

significantly shortened in groups receiving 4mg or 12mg. There was no change in 

refraction, flicker fusion frequency, or pupillary reflex. 

Eye Health 

102

Journal of Clinical Therapeutics & Medicines VOL.22;NO.1;PAGE.41-54(2006) 

The supplementation effect of Astaxanthin on Accommodation and 

Asthenopia 

NAGAKI YASUNORI; MIHARA MIHARU; TSUKAHARA HIROKI; ONO 

SHIGEAKI 

This double blind randomized placebo controlled study examined the 

supplementation effects of Haematococcus (H) pluvialis derived astaxanthin on 

subjects suffering from visual display terminal (VDT) induced visual fatigue. 

Subjects were divided into two groups: 6 mg astaxanthin treated and placebo 

groups. Furthermore, the safety of astaxanthin intake was simultaneously 

assessed. After the 4 week supplementation period, the groups' visual 

accommodation was evaluated as well as a subjective questionnaire designed to 

evaluate visual asthenopia (eye fatigue). Twenty five subjects of the astaxanthin 

treated group and 23 subjects of the placebo group were examined for eye fatigue. 

For safety evaluation, 31 treated subjects and 28 placebo subjects were analysed. 

We report the following observations: 1. In the astaxanthin treated group, the 

change of accommodation before and after supplementation significantly 

improved compared with the placebo group. 2. The astaxanthin supplemented 

group exhibited a significant rate of change in the accommodation compared with 

the placebo group. 3. The subjective questionnaire evaluating visual asthenopia 

revealed a marked reduction in "heavy head" claims. Other typical improvements 

of fatigue symptoms included "dimness of sight" and "stiff shoulders and back". 

4. No significant differences were detected between the treatment and the placebo 

groups after 4 weeks of supplementation in the safety parameters analyzed, and 

adverse event. These results suggest that 6 mg of astaxanthin per day from a H. 

pluvialis algal extract can improve eye fatigue. Moreover, astaxanthin can be 

safely consumed at this level by healthy adults. 

Eye Health 

103


Effect of Astaxanthin on Accommodation and Asthenopia-Efficacy- 

Identification Study in Healthy Volunteers- 

SHIRATORI KENJI; OGAMI KAZUHIRO; NITTA TAKUYA; SHINMEI 

YASUHIRO; CHIN SHINKI; YOSHIDA KAZUHIKO; TSUKAHARA 

HIROKI; TAKEHARA ISAO; ONO SHIGEAKI 

A double-blind study was conducted to confirm the efficacy of H. pluvialis 

Astaxanthin on accommodation and asthenopia and its safety. Two groups of 

subjects were compared, wherein one was given 0mg of Astaxanthin (as a control 

group) and the other was given 6mg of Astaxanthin (AX group). The subjects 

were healthy volunteers who complained of asthenopia. Twenty were enrolled in 

each group, and the testing food was administered during 4 weeks. Sub-objective 

accommodation power, positive accommodation time and negative 

accommodation time were measured before and after administration to 

objectively evaluate the degree of asthenopia. Additionally, subjective degree of 

asthenopia by volunteers was evaluated using VAS. The safety was assessed by 

changes in value of laboratory tests between pre- and post-administrations and by 

the doctor's questions. 1) Sub-objective accommodation power (rate of change) of 

the AX group was significantly higher than that of the control group. 2) The AX 

group showed significantly higher rate of positive and negative accommodation 

times (rate of change) compared to those of the control group. 3) In the AX group, 

subjective degree of asthenopia measured by VAS showed significant 

improvement in two parameters, i.e., "blear-eye feeling" and "tendency of 

irritation" than the control group. 4) No changes in laboratory tests of clinically 

controversial were noted and also no adverse events suggesting causal 

relationship with the testing food were found. In conclusion, administration of 

6mg/day (in a daily dosage of 2 capsules; 3mg/capsule) of H. pluvialis 

Astaxanthin improved accommodation power and subjective symptoms of 

asthenopia. Also, Astaxanthin was confirmed to be completely safe. 

Eye Health 

104

J Pharm Pharmacol. 2008 Oct;60(10):1365-74. 

Astaxanthin, a dietary carotenoid, protects retinal cells against 

oxidative stress in-vitro and in mice in-vivo. 

Nakajima Y, Inokuchi Y, Shimazawa M, Otsubo K, Ishibashi T, Hara H. 

Department of Biofunctional Evaluation, Molecular Pharmacology, Gifu 

Pharmaceutical University, 5-6-1 Mitahora-higashi, Gifu 502-8585, Japan. 

We have investigated whether astaxanthin exerted neuroprotective effects in 

retinal ganglion cells in-vitro and in-vivo. In-vitro, retinal damage was induced by 

24-h hydrogen peroxide (H2O2) exposure or serum deprivation, and cell viability 

was measured using a WST assay. In cultured retinal ganglion cells (RGC-5, a rat 

ganglion cell-line transformed using E1A virus), astaxanthin inhibited the 

neurotoxicity induced by H2O2 or serum deprivation, and reduced the 

intracellular oxidation induced by various reactive oxygen species (ROS). 

Furthermore, astaxanthin decreased the radical generation induced by serum 

deprivation in RGC-5. In mice in-vivo, astaxanthin (100 mg kg(-1), p.o., four 

times) reduced the retinal damage (a decrease in retinal ganglion cells and in 

thickness of inner plexiform layer) induced by intravitreal N-methyl-D-aspartate 

(NMDA) injection. Furthermore, astaxanthin reduced the expressions of 4- 

hydroxy-2-nonenal (4-HNE)-modified protein (indicator of lipid peroxidation) 

and 8-hydroxy-deoxyguanosine (8-OHdG; indicator of oxidative DNA damage). 

These findings indicated that astaxanthin had neuroprotective effects against 

retinal damage in-vitro and in-vivo, and that its protective effects may have been 

partly mediated via its antioxidant effects. 


Eye Health 

105

Invest Ophthalmol Vis Sci. 2008 Apr;49(4):1679-85. 

Inhibition of choroidal neovascularization with an antiinflammatory 

carotenoid astaxanthin. 

Izumi-Nagai K, Nagai N, Ohgami K, Satofuka S, Ozawa Y, Tsubota K, Ohno 

S, Oike Y, Ishida S. 

Laboratory of Retinal Cell Biology, Keio University of Medicne, Tokyo, Japan. 

PURPOSE: Astaxanthin (AST) is a carotenoid found in marine animals and 

vegetables. The purpose of the present study was to investigate the effect of AST 

on the development of experimental choroidal neovascularization (CNV) with 

underlying cellular and molecular mechanisms. METHODS: Laser 

photocoagulation was used to induce CNV in C57BL/6J mice. Mice were 

pretreated with intraperitoneal injections of AST daily for 3 days before 

photocoagulation, and treatments were continued daily until the end of the study. 

CNV response was analyzed by volumetric measurements 1 week after laser 

injury. Retinal pigment epithelium-choroid levels of IkappaB-alpha, intercellular 

adhesion molecule (ICAM)-1, monocyte chemotactic protein (MCP)-1, 

interleukin (IL)-6, vascular endothelial growth factor (VEGF), VEGF receptor 

(VEGFR)-1, and VEGFR-2 were examined by Western blotting or ELISA. AST 

was applied to capillary endothelial (b-End3) cells, macrophages, and RPE cells 

to analyze the activation of NF-kappaB and the expression of inflammatory 

molecules. RESULTS: The index of CNV volume was significantly suppressed 

by treatment with AST compared with that in vehicle-treated animals. AST 

treatment led to significant inhibition of macrophage infiltration into CNV and of 

the in vivo and in vitro expression of inflammation-related molecules, including 

VEGF, IL-6, ICAM-1, MCP-1, VEGFR-1, and VEGFR-2. Importantly, AST 

suppressed the activation of the NF-kappaB pathway, including IkappaB-alpha 

degradation and p65 nuclear translocation. CONCLUSIONS: AST treatment, 

together with inflammatory processes including NF-kappaB activation, 

subsequent upregulation of inflammatory molecules, and macrophage infiltration, 

led to significant suppression of CNV development. The present study suggests 

the possibility of AST supplementation as a therapeutic strategy to suppress CNV 

associated with AMD. 



Eye Health 

106

Nippon Ganka Gakkai Zasshi. 2009 Mar;113(3):403-22; discussion 423. 

[Lifestyle-related diseases and anti-aging ophthalmology: 

suppression of retinal and choroidal pathologies by inhibiting reninangiotensin 

system and inflammation] 

[Article in Japanese] 

Ishida S. 

Inaida Endowed Department of Anti-Aging Ophthalmology, Laboratory of 

Retinal Cell Biology, Center for Integrated Medical Research, Keio University 

School of Medicine, Tokyo, Japan. ishidasu@sc.itc.keio.ac.jp 

Lifestyle-related diseases cause macro-and microangiopathies in the major organs 

including the brain, heart, kidney, and eye, and as a result, shorten the lifespan. The 

renin-angiotensin system (RAS) has recently been shown to contribute to the processes of 

accelerated aging caused by lifestyle-related diseases from visceral obesity in the early 

stage to late-onset organ damage. Vision-threatening diabetic retinopathy and age-related 

macular degeneration (AMD), associated with lifestyle-related diseases as risk factors for 

progression, develop retinal and choroidal neovascularization (CNV), respectively, in 

their advanced stages. We have found that tissue RAS is activated in the pathogenesis of 

diabetic retinopathy and CNV, leading to angiotensin type 1 receptor(AT1-R)-mediated 

expression of inflammation-related molecules including vascular endothelial growth 

factor (VEGF), intercellular adhesion molecule (ICAM)-1, and monocyte chemotactic 

protein(MCP)-1. Neuronal dysfunction in diabetic retinopathy is also shown to result 

from AT1-R-mediated degradation of synaptic proteins. Moreover, we revealed for the 

first time that the receptor for prorenin [(pro) renin receptor] is expressed in the eye, 

although prorenin was until recently believed to be just an inactive precursor of renin. 

Prorenin binds to the receptor that causes dual activation of its intracellular signaling and 

tissue RAS, and this pathogenic mechanism is termed receptor-associated prorenin 

system (RAPS)'. We have demonstrated the contribution of RAPS to the pathogenesis of 

CNV and dual regulation of VEGF and MCP-1 by signal transduction via (pro) renin 

receptor and AT1-R. Next, we report the potential validity of food factor supplements as 

a therapeutic strategy for preventing the retinal and choroidal pathologies driven by RASinduced 

inflammatory and angiogenic molecules. Functional food factors examined 

include lutein in yellow-green vegetables, the omega-3 polyunsaturated fatty acid 

eicosapentaenoic acid purified from fish oil, and red pigment astaxanthin from salmon 

and shrimp. We recently revealed that these food factors prevent intraocular angiogenesis 

and inflammation by inhibiting the expression of inflammatory molecules including 

VEGF, ICAM-1, and MCP-1. Preventive medicine for AMD and diabetic retinopathy, 

both of which have lifestyle-related diseases as a systemic background, has attracted 

growing attention. In the present review, we provide biological evidence for RAS 

inhibition and food factor supplementation in the early intervention for retinal and 

choroidal pathologies as an 'anti-aging ophthalmology' approach. 

Eye Health 

107

Chem Res Toxicol. 2009 Feb 4. [Epub ahead of print] 

Astaxanthin Interacts with Selenite and Attenuates Selenite-Induced 

Cataractogenesis. 

Liao JH, Chen CS, Maher TJ, Liu CY, Lin MH, Wu TH, Wu SH. 

Institute of Biological Chemistry, Academia Sinica, Taipei 115, Taiwan, 

Department of Pharmaceutical Sciences, Massachusetts College of Pharmacy and 

Health Sciences, Boston, Massachusetts 02115, USA, and School of Pharmacy, 

College of Pharmacy, Taipei Medical University, Taipei 110, Taiwan. 

Selenite, the most commonly encountered toxic form of selenium, in overdose, is 

used to induce cataracts in rats. This study demonstrated that selenite, but not 

selenate, would interact with the carotenoid astaxanthin (ASTX), as determined 

using isothermal titration calorimetry and NMR. The maximum absorption of 

ASTX decreased with increasing selenite concentration, indicating that the 

conjugated system of ASTX was changed by selenite. Such interactions between 

ASTX and selenite were also supported by the attenuation of selenite-induced 

turbidity by ASTX (0-12.5 muM) in vitro. In vivo experiments also showed that 

ASTX attenuated selenite-induced cataractogenesis in rats. In summary, this is the 

first report of a direct interaction of ASTX with selenite. This interaction is 

supported by an in vitro assay and may be partially responsible for the ASTX 

observed in vivo protection against selenite-induced cataractogenesis. 


Eye Health 

108

Ophthalmology. 2008 Feb;115(2):324-333.e2. Epub 2007 Aug 22. 

Carotenoids and antioxidants in age-related maculopathy italian study: multifocal 

electroretinogram modifications after 1 year. 

Parisi V, Tedeschi M, Gallinaro G, Varano M, Saviano S, Piermarocchi S; CARMIS 

Study Group. 

Fondazione G. B. Bietti-Istituto di Ricovero e Cura a Carattere Scientifico, Roma, Italy. 

vparisi@tin.it 

OBJECTIVE: To evaluate the influence of short-term carotenoid and antioxidant 

supplementation on retinal function in nonadvanced age-related macular degeneration 

(AMD). DESIGN: Randomized controlled trial. PARTICIPANTS: Twenty-seven 

patients with nonadvanced AMD and visual acuity > or =0.2 logarithm of the minimum 

angle of resolution were enrolled and randomly divided into 2 age-similar groups: 15 

patients had oral supplementation of vitamin C (180 mg), vitamin E (30 mg), zinc (22.5 

mg), copper (1 mg), lutein (10 mg), zeaxanthin (1 mg), and astaxanthin (4 mg) (AZYR 

SIFI, Catania, Italy) daily for 12 months (treated AMD [T-AMD] group; mean age, 

69.4+/-4.31 years; 15 eyes); 12 patients had no dietary supplementation during the same 

period (nontreated AMD [NT-AMD] group; mean age, 69.7+/-6.23 years; 12 eyes). At 

baseline, they were compared with 15 age-similar healthy controls. METHODS: 

Multifocal electroretinograms in response to 61 M-stimuli presented to the central 20 

degrees of the visual field were assessed in pretreatment (baseline) conditions and, in 

nonadvanced AMD patients, after 6 and 12 months. MAIN OUTCOME MEASURES: 

Multifocal electroretinogram response amplitude densities (RAD, nanovolt/deg(2)) of the 

N1-P1 component of first-order binary kernels measured from 5 retinal eccentricity areas 

between the fovea and midperiphery: 0 degrees to 2.5 degrees (R1), 2.5 degrees to 5 

degrees (R2), 5 degrees to 10 degrees (R3), 10 degrees to 15 degrees (R4), and 15 

degrees to 20 degrees (R5). RESULTS: At baseline, we observed highly significant 

reductions of N1-P1 RADs of R1 and R2 in T-AMD and NT-AMD patients when 

compared with healthy controls (1-way analysis of variance P0.05) from 

controls. No significant differences (P>0.05) were observed in N1-P1 RADs of R1-R5 

between T-AMD and NT-AMD at baseline. After 6 and 12 months of treatment, T-AMD 

eyes showed highly significant increases in N1-P1 RADs of R1 and R2 (P0.05) change was observed in N1-P1 RADs of R3-R5. No 

significant (P>0.05) changes were found in N1-P1 RADs of R1-R5 in NT-AMD eyes. 

CONCLUSIONS: In nonadvanced AMD eyes, a selective dysfunction in the central 

retina (0 degrees -5 degrees ) can be improved by the supplementation with carotenoids 

and antioxidants. No functional changes are present in the more peripheral (5 degrees -20 

degrees ) retinal areas. 


Eye Health 

109

J Photochem Photobiol B. 2007 Jul 27;88(1):1-10. Epub 2007 May 1. 

Lutein, zeaxanthin and astaxanthin protect against DNA damage in 

SK-N-SH human neuroblastoma cells induced by reactive nitrogen 

species. 

Santocono M, Zurria M, Berrettini M, Fedeli D, Falcioni G. 

Medical Department, SIFI SpA, Via E. Patti 36, Lavinaio (CT), Italy. 

The purpose of this study was to evaluate the ability of the predominant 

carotenoids (lutein and zeaxanthin) of the macular pigment of the human retina, to 

protect SK-N-SH human neuroblastoma cells against DNA damage induced by 

different RNOS donors. Although astaxanthin has never been isolated from the 

human eye, it was included in this study because its structure is very close to that 

of lutein and zeaxanthin and because it affords protection from UV-light. DNA 

damage was induced by GSNO-MEE, a nitric oxide donor, by Na(2)N(2)O(3), a 

nitroxyl anion donor and by SIN-1, a peroxynitrite-generating agent. DNA 

damage was assessed using the comet assay, a rapid and sensitive single cell gel 

electrophoresis technique able to detect primary DNA damage in individual cells. 

The tail moment parameter was used as an index of DNA damage. The values of 

tail moment increased in all the samples incubated with the RNOS donors, 

indicating DNA impairment. Data obtained show that the ability of zeaxanthin, 

lutein, and astaxanthin to reduce the DNA damage depends on the type of RNOS 

donor and the carotenoid concentration used. All the carotenoids studied were 

capable of protecting against DNA damage in neuroblastoma cells when the cells 

were exposed to GSNO-MEE. However, a different behaviour was present when 

the other two RNOS donors were used. The presence of a carotenoid alone 

(without an RNOS donor) did not cause DNA damage. Spectrophotometric 

studies showed that the order with which tested carotenoids reacted with RNOS 

was not always in agreement with the DNA protection results. The data from this 

study provides additional information on the activities of the macular pigment 

carotenoids of the human retina. 



Eye Health 

110


Astaxanthin protects against oxidative stress and calcium-induced 

porcine lens protein degradation. 




























Eye Health 

111

Exp Eye Res. 2006 Feb;82(2):275-81. Epub 2005 Aug 26. 

Suppressive effects of astaxanthin against rat endotoxin-induced 

uveitis by inhibiting the NF-kappaB signaling pathway. 

Suzuki Y, Ohgami K, Shiratori K, Jin XH, Ilieva I, Koyama Y, Yazawa K, 

Yoshida K, Kase S, Ohno S. 


Graduate School of Medicine, N15 W7, Sapporo 060-8638, Japan. 

We investigated the effects of astaxanthin (AST), a carotenoid, on endotoxininduced 

uveitis (EIU), and over the course of the disease measured the expression 

of inflammatory cytokines and chemokines in the presence or absence of AST. 

EIU was induced in male Lewis rats by footpad injection of lipopolysaccharide 

(LPS). The animals were randomly divided to 12 groups with eight animals in 

each. Immediately after the inoculation, AST (1, 10, or 100 mg kg(-1)) was 

injected intravenously. Aqueous humour was collected at 6, 12 and 24 hr after 

LPS inoculation and the number of infiltrating cells in the anterior chamber was 

counted. In addition, we assayed the concentration of protein, nitric oxide (NO), 

tumour necrosis factor-alpha (TNF-alpha) and prostaglandin E2 (PGE2). 

Immunohistochemical staining with a monoclonal antibody against activated NFkappaB 

was performed in order to evaluate the effects of AST on NF-kappaB 

activation. Rats injected with AST showed a significant decrease in the number of 

infiltrating cells in the anterior chamber and additionally there was a significantly 

lower concentration of protein, NO, TNF-alpha and PGE2 in the aqueous humour. 

Moreover, even early stages of EIU were suppressed by injection of AST. The 

number of activated NF-kappaB-positive cells was lower in iris-ciliary bodies 

treated with 10 or 100 mg kg(-1) AST at 3 hr after LPS injection. These results 

suggest that AST reduces ocular inflammation in eyes with EIU by 

downregulating proinflammatory factors and by inhibiting the NF-kappaBdependent 

signaling pathway. 



Eye Health 

112

J Nutr. 2004 Dec;134(12):3225-32. 

Xanthophylls and alpha-tocopherol decrease UVB-induced lipid 

peroxidation and stress signaling in human lens epithelial cells. 

Chitchumroonchokchai C, Bomser JA, Glamm JE, Failla ML. 

Ohio State University Interdisciplinary PhD Program in Nutrition, Ohio State 

University, Columbus, OH 43210, USA. 

Epidemiological studies suggest that consumption of vegetables rich in the 

xanthophylls lutein (LUT) and zeaxanthin (ZEA) reduces the risk for developing 

age-related cataract, a leading cause of vision loss. Although LUT and ZEA are 

the only dietary carotenoids present in the lens, direct evidence for their 

photoprotective effect in this organ is not available. The present study examined 

the effects of xanthophylls and alpha-tocopherol (alpha-TC) on lipid peroxidation 

and the mitogen-activated stress signaling pathways in human lens epithelial 

(HLE) cells following ultraviolet B light (UVB) irradiation. When presented with 

LUT, ZEA, astaxanthin (AST), and alpha-TC as methyl-beta-cyclodextrin 

complexes, HLE cells accumulated the lipophiles in a concentration- and timedependent 

manner with uptake of LUT exceeding that of ZEA and AST. 

Pretreatment of cultures with either 2 micromol/L xanthophyll or 10 micromol/L 

alpha-TC for 4 h before exposure to 300 J/m(2) UVB radiation decreased lipid 

peroxidation by 47-57% compared with UVB-treated control HLE cells. 

Pretreatment with the xanthophylls and alpha-TC also inhibited UVB-induced 

activation of c-JUN NH(2)-terminal kinase (JNK) and p38 by 50-60 and 25-32%, 

respectively. There was substantial inhibition of UVB-induced JNK and p38 

activation for cells containing 2.3 nmol alpha-TC/mg protein was 

required to significantly decrease UVB-induced stress signaling. These data 

suggest that xanthophylls are more potent than alpha-TC for protecting human 

lens epithelial cells against UVB insult. 



Eye Health 

113

Cataract formation in Atlantic salmon, Salmo salar L., smolt 

relative to dietary pro- and antioxidants and lipid level. 

Waagbø R, Hamre K, Bjerkås E, Berge R, Wathne E, Lie O, Torstensen B. 

National Institute of Nutrition and Seafood Research, Bergen, Norway. 

rune.waagbo@nutr.fiskeridir.no 

The development of cataracts in Atlantic salmon, Salmo salar L., was studied in 

16 groups of smolts fed diets differing in prooxidant (iron, copper, manganese) 

and antioxidant (vitamin E, vitamin C, astaxanthin) composition and lipid level 

for 23 weeks in sea water, using a 2(7-3) reduced factorial design. The seven 

dietary variables were systematically varied at low (requirement level and 150 g 

lipid kg(-1)) and high levels (below known toxic levels and 320 g lipid kg(-1)). A 

mean endpoint cataract incidence of approximately 36% was observed. High 

dietary levels of vitamin C and astaxanthin reduced cataract frequency, whereas 

high dietary lipid level, iron and manganese were associated with increased 

cataract frequencies. Considering the nutritional status of selected organs of the 

fish, only the status of ascorbic acid correlated negatively to cataract development 

(P < 0.05). The lens glutathione (GSH) status was not correlated to cataract 

frequency, nor statistically explained by the dietary variables. However, the study 

shows that balancing the diet with respect to pro- and antioxidant nutrients may 

significantly protect Atlantic salmon against development of cataracts. An 

incidence of reversible osmotic cataract observed at week 14 was positively 

correlated to plasma glucose concentration. 



Eye Health 

114


Effects of Astaxanthin on Accommodative Recovery 

TAKAHASHI NANAKO (Kajitaganka) KAJITA MASAYOSHI (Kajitaganka) 

Effects of astaxanthin on accommodative recovery derived from a rest after VDT 

(visual display terminal) working was studied. Ten healthy volunteers were 

entered into the study, and except one subject who developed allergic 

conjunctivitis during the study, 9 of whom were evaluated (9 dominant eyes) by 

values of objective diopter, HFC (High Frequency Component in Accommodative 

micro-fluctuation) and accommodative reaction. Consequently, increase of HFC 

after the rest was significantly restrained by astaxanthin uptake compared to that 

shortly after working. Therefore, Astaxanthin was suggested to have effects on 

accommodation during recovery process of accommodative fatigue to relieve 

fatigue rapidly. 

Eye Health 

115

Journal of Traditional Medicines VOL.19;NO.5;PAGE.170-173(2002) 

Effects of astaxanthin on accommodation, critical flicker fusion, and 

pattern visual evoked potential in visual display terminal workers. 

NAGAKI Y; HAYASAKA S ; YAMADA T ; HAYASAKA Y; SANADA M; 

UONOMI T 

We evaluated the effects of astaxanthin, a red carotenoid, on accommodation, 

critical flicker fusion(CFF), and pattern visual evoked potential(PVEP) in visual 

display terminal(VDT) workers. As controls, 13 non-VDT workers received no 

supplementation (Group A). Twenty-six VDT workers were randomized into 2 

groups: Group B consisted of 13 subjects who received oral astaxanthin, 5mg/day, 

for 4 weeks, and Group C consisted of 13 subjects who received an oral placebo, 

5mg/day, for 4 weeks. No significant difference in age was noted among the 3 

groups. A double-masked study was designed in Groups B and C. 

Accommodation amplitude in Group A was 3.7.+-.1.5 diopters. Accommodation 

amplitudes (2.3.+-.1.4 and 2.2.+-.1.0 diopters) in Groups B and C before 

supplementation were significantly (p


Effects of Astaxanthin on Accommodation and Asthenopia-Dose 

Finding Study in Healthy Volunteers- 

NITTA TAKUYA; OGAMI KAZUHIRO; SHIRATORI KENJI; SHINMEI 

YASUHIRO; CHIN SHINKI; YOSHIDA KAZUHIKO; TSUKAHARA 

HIROKI; ONO SHIGEAKI 

A double-blind study was conducted in healthy volunteers to objectively evaluate 

the optimum dose and safety of astaxanthin (AX) on accommodation and 

asthenopia. The subjects were divided into 3 groups: 0mg (AX 0mg group), 6mg 

(AX 6mg group) and 12mg (AX 12mg group) of astaxanthin administered. Ten 

subjects, total thirty subjects were included in each group. Mean time consumed 

for close working (e.g., VDT working) was approximately 7 hours a day. The 

testing food was given to the subjects for 4 weeks. Then, the subjects were traced 

for 4 weeks and assessed by comparison of the observed values between pre- and 

post-dosing. As a result 1. Objective accommodation power of the AX 12mg 

group was significantly increased compared to that of pre-dosing. 2. Positive 

accommodation time was significantly shortened in the AX 6mg and the 12mg 

groups compared to those of pre-dosing, and negative accommodation time was 

significantly shortened in the AX 0mg and the 6mg groups compared to those of 

pre-dosing. 3. According to the assessment by VAS, many parameters in 

subjective symptoms were improved in the AX 6mg group. 4. No changes were 

noted in laboratory tests of controversial in clinical setting due to AX uptake. 

Also, there were no adverse events caused by the administration of the testing 

food. In conclusion, accommodation power and subjective symptoms relating 

asthenopia were improved by taking 6mg/day of astaxanthin, therefore more than 

6mg/day was considered to be optimal dosage of astaxanthin. 

Eye Health 

117

Bechettobyo ni kansuru Chosa Kenkyu Heisei 14 Nendo Sokatsu, Buntan Kenkyu 

Hokokusho VOL.;NO.;PAGE.98-99(2003) 

Research on the anti-inflammatory effect of astaxanthin 

ONO SHIGEAKI; OGAMI KAZUHIRO; SHIRATORI KENJI; ILIEVA I; 

KOTAKE SATOSHI; NISHIDA TOMOMI; MIZUKI NOBUHISA 

The effect of astaxanthin (AST) was examined in rat model of the endotoxin 

induced uveitis. As the result, the protein concentration in the hydatoid lowered 

obviously in the group which administered 10 (AST10) or 100mg/kg (AST100) of 

AST in comparison with control animals.The number of inflammatory cells was 

significantly decreased only in AST100 group. The effect of AST on protein 

concentration and cell numbers in the hydatoid in AST100 group was almost 

equivalent to those of 10mg/kg of prednisolone (PSL) administrated group. Any 

side effects by AST administration could not be observed. AST showed dosedependent 

inhibitory effect in this model. Therefore, it was indicated that AST 

could be utilized as a new antiphlogistic for ophthalmia disease. 

Eye Health 

118

Atarashii Ganka, 25(10):1461-1464 (In Japanese). 2008 

Intraocular penetration of astaxanthin in rabbit eyes 

Fukuda et al., 

In a new study, natural astaxanthin extract derived from Haematococcus 

microalgae was detected in the iris/ciliary body of New Zealand Albino (NZW) 

Rabbit Eyes 24 hours after ingestion. 

Astaxanthin has been reported to have many benefits in the eye. Several human 

clinical studies reported the alleviation of eye fatigue (by improving 

accommodation function) in visual display terminal (VDT) workers after oral 

supplementation. However, up to now there has been no intraocular kinetic 

information available. In collaboration between the Ophthalmology Department 

of Kanazawa Medical University, Japan, and Fuji Chemical Industry, Japan, 

researchers investigated the ocular and blood serum levels of astaxanthin in 24 

NZW albino rabbits. After administering a 100 mg/kg single oral dose, 

astaxanthin was determined by careful extraction followed by HPLC analysis over 

a period of 168 hours. According to the astaxanthin detection system, the time 

taken to reach maximum presence (Tmax) in serum and iris/ciliary body was 9 

hours (at Cmax 61.3 ng/mL) and 24 hours (at Cmax 79.3) respectively. In other 

human studies with oral intake of astaxanthin, the Tmax in serum ranged between 

9 and 12 hours. 

The intraocular penetration kinetics could have a similar pattern to humans but 

further study is necessary. This study adds to the growing body of science 

supporting astaxanthin’s benefits for eye fatigue caused by VDT use. 

Eye Health 

119

Journal of the Eye VOL.23;NO.6;PAGE.829-834(2006) 

Effects of Astaxanthin on Eyestrain Induced by Accommodative 

Dysfunction 

IWASAKI TSUNETO; TAHARA AKIHIKO 

We investigated effects of astaxanthin on eyestrain induced by accommodative 

dysfunction. The 10 healthy subjects received 6mg/day of astaxanthin (Ax group) 

or 0mg/day (placebo; P group) for 14 days, and were then assigned a near visual 

task for 20min. Accommodative function and subjective symptoms relating to 

eyestrain were measured before and after the task, and after the 10-minute rest 

following the task. The data were then compared between Ax and P groups by the 

double-blind cross-over method. After the task, accommodation contraction and 

relaxation times were extended in both the Ax and P groups. Comparison between 

the two groups showed that after the task, accommodation relaxation time was 

significantly extended in P group, in contrast to Ax. Accommodative contraction 

and relaxation times were significantly prolonged after the 10-minute rest in P 

group as compared to Ax. The symptoms eye fatigue, eye heaviness, blurred 

vision and eye dryness in P group were increased, but Ax group showed increased 

in eye fatigue and eye heaviness. On the basis of these results, we concluded that 

astaxanthin has the effects of reducing and preventing eyestrain induced by 

accommodative dysfunction. 

Eye Health 

120

Journal of Traditional Medicines 2002: 19 (5), 170 – 173. 

Effects of Astaxanthin on accommodation, critical flicker fusion, and pattern visual 

evoked potential in visual display terminal workers. 

Nagaki Y., Hayasaka S., Yamada T., Hayasaka Y., Sanada M., Uonomi T. 

Working for long periods at visual display terminals reportedly induces various 

visual problems such as eye strain, blurring and diplopia (a disorder of vision in 

which two images of a single object are seen because of unequal action of the eye 

muscles – also called double vision). In a double blind study performed in Japan, 

after four weeks of supplementation with 5 mg of Astaxanthin per day (extracted 

from Haematococcus Pluvialis algae meal) the authors reported a 46% reduction 

of eye strain subjects and higher accommodation amplitude in visual display 

terminal subjects. 

Although the mechanism of action is unclear, Astaxanthin’s potent antioxidant 

properties may relieve chronic stress of visual display terminal use that may 

induce hypofunction of the ciliary body, resulting in decreased accommodation. 

Eye Health 

121

Invest Ophthalmol Vis Sci. 2003 Jun;44(6):2694-701 

Effects of astaxanthin on lipopolysaccharide-induced inflammation 

in vitro and in vivo 

Ohgami K, Shiratori K, Kotake S, Nishida T, Mizuki N, Yazawa K, Ohno S 


Graduate School of Medicine, Sapporo, Japan. kohgami@med.hokudai.ac.jp 

PURPOSE: Astaxanthin (AST) is a carotenoid that is found in marine animals and 

vegetables. Several previous studies have demonstrated that AST exhibits a wide 

variety of biological activities including antioxidant, antitumor, and anti- 

Helicobacter pylori effects. In this study, attention was focused on the antioxidant 

effect of AST. The object of the present study was to investigate the efficacy of 

AST in endotoxin-induced uveitis (EIU) in rats. In addition, the effect of AST on 

endotoxin-induced nitric oxide (NO), prostaglandin E2 (PGE2), and tumor 

necrosis factor (TNF)-alpha production in a mouse macrophage cell line (RAW 

264.7) was studied in vitro. METHODS: EIU was induced in male Lewis rats by a 

footpad injection of lipopolysaccharide (LPS). AST or prednisolone was 

administered intravenously at 30 minutes before, at the same time as, or at 30 

minutes after LPS treatment. The number of infiltrating cells and protein 

concentration in the aqueous humor collected at 24 hours after LPS treatment was 

determined. RAW 264.7 cells were pretreated with various concentrations of AST 

for 24 hours and subsequently stimulated with 10 microg/mL of LPS for 24 hours. 

The levels of PGE2, TNF-alpha, and NO production were determined in vivo and 

in vitro. RESULTS: AST suppressed the development of EIU in a dose-dependent 

fashion. The anti-inflammatory effect of 100 mg/kg AST was as strong as that of 

10 mg/kg prednisolone. AST also decreased production of NO, activity of 

inducible nitric oxide synthase (NOS), and production of PGE2 and TNF-alpha in 

RAW264.7 cells in vitro in a dose-dependent manner. CONCLUSIONS: This 

study suggests that AST has a dose-dependent ocular anti-inflammatory effect, by 

the suppression of NO, PGE2, and TNF-alpha production, through directly 

blocking NOS enzyme activity. 

Eye Health 

122


The Effect of Astaxanthin on Retinal Capillary Blood Flow in 

Normal Volunteers 

NAGAKI YASUNORI; MIHARA MIHARU; TAKAHASHI JIRO; KITAMURA 

AKITOSHI; HORITA YOSHIHARU; SUGIURA YURI; TSUKAHARA 

HIROKI 

Objective: We evaluated the effect of astaxanthin on retinal circulation in healthy 

volunteers. Design A double blind randomized placebo controlled study. 

Methods: Thirty-six volunteers were randomized into two groups: Astaxanthin 

group that consisted of 18 subjects who received oral astaxanthin, 6mg/day, for 4 

weeks and a placebo group that consisted of 18 subjects who received an identical 

looking oral placebo for 4 weeks. Retinal capillary blood flow was measured by 

the Heidelberg Retina Flowmeter. Changes in blood pressure, blood cell counts, 

fasting plasma glucose level, fasting plasma astaxanthin level, retinal capillary 

blood flow, intraocular pressure, inquiry about eye strain were examined before 

and after supplementation in both groups. Results: The fasting plasma astaxanthin 

level in the astaxanthin group was significantly (P


Sports Performance Benefits from Taking Natural Astaxanthin 

Characterized by Visual Acuity and Muscle Fatigue Improvement in 

Humans. 

SAWAKI KEISUKE; YOSHIGI HIROSHI; AOKI KAZUHIRO; KOIKAWA 

NATSUE; AZUMANE AKITO; KANEKO KESATOKI; YAMAGUCHI 

MASAHIRO 

The effects of astaxanthin on visual acuity and muscle fatigue were studied. 

Astaxanthin (3,3'-Dihydroxy-.BETA.,.BETA.-carotene-4,4'-dione) is a red 

pigment found in salmon and krill and has strong antioxidant properties. In the 

two supplementation studies, astaxanthin extracted from algae (Haematococcus 

pluvialis) was used. Four visual acuity parameters were examined in experiment 

A in 18 healthy adult male volunteers that were equally divided into two groups 

(treatment and control). The measured parameters were deep vision, critical 

flicker fusion, static and kinetic visual acuity before and after supplementation. A 

second investigation (experiment B) involved 16 adult male volunteers to 

establish the effect of astaxanthin supplementation on the build up of lactic acid 

before and after running 1200 metres. In both experiments, the treated groups 

ingested an astaxanthin capsule per day for 4 weeks (6mg astaxanthin per day) 

and the control groups received a placebo capsule. Results: In experiment A, the 

deep vision and the critical flicker fusion of the treated groups were significantly 

improved compared to the control group. No effects of treated group were 

observed on static and kinetic visual acuity. In experiment B, serum lactic acid 

concentration at 2 minutes after activity (1,200m running) of the treatment group 

was significantly lower than that of the control one. No other effects related to 

supplementation of astaxanthin on serum biological and hematological 

examinations were observed. Based on these preliminary findings, it suggested 

that supplementation of astaxanthin is effective for the improvement of visual 

acuity and muscle fatigue that may lead to sports performance benefits. 

Eye Health 

124

Regul Toxicol Pharmacol. 2010 Oct;58(1):121-30. Epub 2010 May 8. 

Suppressive effect of astaxanthin on retinal injury induced by elevated 

intraocular pressure. 

Cort A, Ozturk N, Akpinar D, Unal M, Yucel G, Ciftcioglu A, Yargicoglu P, Aslan M. 

Department of Biochemistry, Akdeniz University, School of Medicine, Antalya 07070, 

Turkey. 


The aim of this study was to clarify the possible protective effect of astaxanthin (ASX) 

on the retina in rats with elevated intraocular pressure (EIOP). Rats were randomly 

divided into two groups which received olive oil or 5mg/kg/day ASX for a period of 8 

weeks. Elevated intraocular pressure was induced by unilaterally cauterizing three 

episcleral vessels and the unoperated eye served as control. At the end of the 

experimental period, neuroprotective effect of ASX was determined via 

electrophysiological measurements of visual evoked potentials (VEP) and rats were 

subsequently sacrificed to obtain enucleated globes which were divided into four groups 

including control, ASX treated, EIOP, EIOP+ASX treated. Retinoprotective properties of 

ASX were determined by evaluating retinal apoptosis, protein carbonyl levels and nitric 

oxide synthase-2 (NOS-2) expression. Latencies of all VEP components were 

significantly prolonged in EIOP and returned to control levels following ASX 

administration. When compared to controls, EIOP significantly increased retinal protein 

oxidation which returned to baseline levels in ASX treated EIOP group. NOS-2 

expression determined by Western blot analysis and immunohistochemical staining was 

significantly greater in rats with EIOP compared to ASX and control groups. Retinal 

TUNEL staining showed apoptosis in all EIOP groups; however ASX treatment 

significantly decreased the percent of apoptotic cells when compared to non treated 

ocular hypertensive controls. The presented data confirm the role of oxidative injury in 

EIOP and highlight the protective effect of ASX in ocular hypertension. 

Eye Health 

125


Astaxanthin protects against oxidative stress and 

calcium-induced porcine lens protein degradation. 


























PMID: 16536628 [PubMed - indexed for MEDLIN 

Eye Health 

126

J Pharm Pharmacol. 2008 Oct;60(10):1365-74. 

Astaxanthin, a dietary carotenoid, 

protects retinal cells against oxidative 

stress in-vitro and in mice in-vivo. 

Nakajima Y, Inokuchi Y, Shimazawa M, Otsubo K, Ishibashi T, Hara H. 

Source 

Department of Biofunctional Evaluation, Molecular Pharmacology, Gifu Pharmaceutical 

University, 5-6-1 Mitahora-higashi, Gifu 502-8585, Japan. 


We have investigated whether astaxanthin exerted neuroprotective effects in retinal 

ganglion cells in-vitro and in-vivo. In-vitro, retinal damage was induced by 24-h 

hydrogen peroxide (H2O2) exposure or serum deprivation, and cell viability was 

measured using a WST assay. In cultured retinal ganglion cells (RGC-5, a rat ganglion 

cell-line transformed using E1A virus), astaxanthin inhibited the neurotoxicity induced 

by H2O2 or serum deprivation, and reduced the intracellular oxidation induced by 

various reactive oxygen species (ROS). Furthermore, astaxanthin decreased the radical 

generation induced by serum deprivation in RGC-5. In mice in-vivo, astaxanthin (100 mg 

kg(-1), p.o., four times) reduced the retinal damage (a decrease in retinal ganglion cells 

and in thickness of inner plexiform layer) induced by intravitreal N-methyl-D-aspartate 

(NMDA) injection. Furthermore, astaxanthin reduced the expressions of 4-hydroxy-2- 

nonenal (4-HNE)-modified protein (indicator of lipid peroxidation) and 8-hydroxydeoxyguanosine 

(8-OHdG; indicator of oxidative DNA damage). These findings 

indicated that astaxanthin had neuroprotective effects against retinal damage in-vitro and 

in-vivo, and that its protective effects may have been partly mediated via its antioxidant 

effects. 


Eye Health 

127

Neuroprotective 

Forum Nutr. 2009;61:129-35. Epub 2009 Apr 7. 

Astaxanthin protects neuronal cells against oxidative damage and is 

a potent candidate for brain food. 

Liu X, Osawa T. 

Graduate School of Bioagricultural Science, Nagoya University, Nagoya, Japan. 

Astaxanthin (AST) is a powerful antioxidant that occurs naturally in a wide 

variety of living organisms. Based on the report claiming that AST could cross the 

brain-blood barrier, the aim of this study was to investigate the neuroprotective 

effect of AST by using an oxidative stress-induced neuronal cell damage system. 

The treatment with DHA hydroperoxide (DHA-OOH) or 6-hydroxydopamine (6- 

OHDA), either of which is a reactive oxygen species (ROS)-inducing neurotoxin, 

led to a significant decrease in viable dopaminergic SH-SY5Y cells by the MTT 

assay, whereas a significant protection was shown when the cells were pretreated 

with AST. Moreover, 100 nM AST pretreatment significantly inhibited 

intracellular ROS generation that occurred in either DHA-OOH- or 6-OHDAtreated 

cells. The neuroprotective effect of AST is suggested to be dependent 

upon its antioxidant potential and mitochondria protection; therefore, it is strongly 

suggested that treatment with AST may be effective for oxidative stressassociated 

neurodegeneration and a potential candidate for natural brain food. 



128

FASEB J. 2009 Jun;23(6):1958-68. Epub 2009 Feb 13. 

Astaxanthin reduces ischemic brain injury in adult rats. 

Shen H, Kuo CC, Chou J, Delvolve A, Jackson SN, Post J, Woods AS, Hoffer 

BJ, Wang Y, Harvey BK. 

National Institute on Drug Abuse, NIH, 251 Bayview Blvd., Baltimore, MD 

21224, USA. 

Astaxanthin (ATX) is a dietary carotenoid of crustaceans and fish that contributes 

to their coloration. Dietary ATX is important for development and survival of 

salmonids and crustaceans and has been shown to reduce cardiac ischemic injury 

in rodents. The purpose of this study was to examine whether ATX can protect 

against ischemic injury in the mammalian brain. Adult rats were injected 

intracerebroventricularly with ATX or vehicle prior to a 60-min middle cerebral 

artery occlusion (MCAo). ATX was present in the infarction area at 70-75 min 

after onset of MCAo. Treatment with ATX, compared to vehicle, increased 

locomotor activity in stroke rats and reduced cerebral infarction at 2 d after 

MCAo. To evaluate the protective mechanisms of ATX against stroke, brain 

tissues were assayed for free radical damage, apoptosis, and excitoxicity. ATX 

antagonized ischemia-mediated loss of aconitase activity and reduced glutamate 

release, lipid peroxidation, translocation of cytochrome c, and TUNEL labeling in 

the ischemic cortex. ATX did not alter physiological parameters, such as body 

temperature, brain temperature, cerebral blood flow, blood gases, blood pressure, 

and pH. Collectively, our data suggest that ATX can reduce ischemia-related 

injury in brain tissue through the inhibition of oxidative stress, reduction of 

glutamate release, and antiapoptosis. ATX may be clinically useful for patients 

vulnerable or prone to ischemic events. 



PMCID: PMC2698661 [Available on 2010/06/01] 


129

J Clin Biochem Nutr. 2009 May;44(3):280-4. Epub 2009 Apr 25. 

Preliminary Clinical Evaluation of Toxicity and Efficacy of A New 

Astaxanthin-rich Haematococcus pluvialis Extract. 

Satoh A, Tsuji S, Okada Y, Murakami N, Urami M, Nakagawa K, Ishikura 

M, Katagiri M, Koga Y, Shirasawa T. 

Life Science Institute, Yamaha Motor Co., Ltd., 3001-10 Kuno, Fukuroi, 

Shizuoka 437-0061, Japan. 

Astaxanthin (Ax), a carotenoid ubiquitously distributed in microorganisms, fish, 

and crustaceans, has been known to be a potent antioxidant and hence exhibit 

various physiological effects. We attempted in these studies to evaluate clinical 

toxicity and efficacy of long-term administration of a new Ax product, by 

measuring biochemical and hematological blood parameters and by analyzing 

brain function (using CogHealth and P300 measures). Ax-rich Haematococcus 

pluvialis extracts equivalent to 4, 8, 20 mg of Ax dialcohol were administered to 

73, 38, and 16 healthy adult volunteers, respectively, once daily for 4 weeks to 

evaluate safety. Ten subjects with age-related forgetfulness received an extract 

equivalent to 12 mg in a daily dosing regimen for 12 weeks to evaluate efficacy. 

As a result, no abnormality was observed and efficacy for age-related decline in 

cognitive and psychomotor functions was suggested. 




130

Brain Res. 2009 Feb 13;1254:18-27. Epub 2008 Dec 3. 

Astaxanthin inhibits reactive oxygen species-mediated cellular 

toxicity in dopaminergic SH-SY5Y cells via mitochondria-targeted 

protective mechanism. 

Liu X, Shibata T, Hisaka S, Osawa T. 

Laboratory of Food and Biodynamics, Graduate School of Bioagricultural 

Science, Nagoya University, Furo-cho, Nagoya 464-8601, Japan. 

Astaxanthin is a powerful antioxidant that occurs naturally in a wide variety of 

living organisms. The aim of this study is to investigate the effect and the 

mechanism of astaxanthin on reactive oxygen species (ROS)-mediated apoptosis 

in dopaminergic SH-SY5Y cells. The treatment with DHA hydroperoxide (DHA- 

OOH) or 6-hydroxydopamine (6-OHDA), either of which is ROS-inducing 

neurotoxin, led to a significant decrease in viable dopaminergic SH-SY5Y cells 

by MTT assay, whereas a significant protection was shown while the cells were 

pretreated with astaxanthin. Moreover, 100 nM astaxanthin pretreatment 

significantly inhibited apoptosis, mitochondrial abnormalities and intracellular 

ROS generation occurred in either DHA-OOH- or 6-OHDA-treated cells. The 

neuroprotective effect of astaxanthin is suggested to be dependent upon its 

antioxidant potential and mitochondria protection; therefore, it is suggested that 

astaxanthin may be an effective treatment for oxidative stress-associated 

neurodegeneration. 



131

J Neurochem. 2008 Dec;107(6):1730-40. Epub 2008 Nov 7. 

Protective effects of astaxanthin on 6-hydroxydopamine-induced 

apoptosis in human neuroblastoma SH-SY5Y cells. 

Ikeda Y, Tsuji S, Satoh A, Ishikura M, Shirasawa T, Shimizu T. 

Research Team for Molecular Biomarkers, Tokyo Metropolitan Institute of 

Gerontology, Tokyo, Japan. 

Parkinson's disease (PD) is a neurodegenerative disorder characterized by 

selective loss of dopaminergic neurons in the substantia nigra pars compacta. 

Although understanding of the pathogenesis of PD remains incomplete, increasing 

evidence from human and animal studies has suggested that oxidative stress is an 

important mediator in its pathogenesis. Astaxanthin (Asx), a potent antioxidant, 

has been thought to provide health benefits by decreasing the risk of oxidative 

stress-related diseases. This study examined the protective effects of Asx on 6- 

hydroxydopamine (6-OHDA)-induced apoptosis in the human neuroblastoma cell 

line SH-SY5Y. Pre-treatment of SH-SY5Y cells with Asx suppressed 6-OHDAinduced 

apoptosis in a dose-dependent manner. In addition, Asx strikingly 

inhibited 6-OHDA-induced mitochondrial dysfunctions, including lowered 

membrane potential and the cleavage of caspase 9, caspase 3, and poly(ADPribose) 

polymerase. In western blot analysis, 6-OHDA activated p38 MAPK, c- 

jun NH(2)-terminal kinase 1/2, and extracellular signal-regulated kinase 1/2, 

while Asx blocked the phosphorylation of p38 MAPK but not c-jun NH(2)- 

terminal kinase 1/2 and extracellular signal-regulated kinase 1/2. Pharmacological 

approaches showed that the activation of p38 MAPK has a critical role in 6- 

OHDA-induced mitochondrial dysfunctions and apoptosis. Furthermore, Asx 

markedly abolished 6-OHDA-induced reactive oxygen species generation, which 

resulted in the blockade of p38 MAPK activation and apoptosis induced by 6- 

OHDA treatment. Taken together, the present results indicated that the protective 

effects of Asx on apoptosis in SH-SY5Y cells may be, at least in part, attributable 

to the its potent antioxidative ability. 




132

Alcohol Clin Exp Res. 2008 Aug;32(8):1417-21. Epub 2008 Jun 6. 

Dose-dependent effects of astaxanthin on cortical spreading 

depression in chronically ethanol-treated adult rats. 

Abadie-Guedes R, Santos SD, Cahú TB, Guedes RC, de Souza Bezerra R. 

Departamento de Bioquímica, Universidade Federal de Pernambuco, 50670-901 

Recife, PE, Brazil. 

BACKGROUND: The consumption of alcoholic drinks is a frequent drug-abuse 

situation, which is associated to a wide variety of pathological disturbances 

affecting several organs, including the brain. We have previously shown in the 

developing rat brain that ethanol intake facilitates the propagation of cortical 

spreading depression (CSD), an excitability-related neural phenomenon present in 

several animal species. This electrophysiological effect was attenuated by a 

shrimp (Litopenaeus vannamei) carotenoids extract. Here we investigated the 

effects of pure astaxanthin, the main carotenoid found in shrimp, on CSD. 

METHODS: Adult Wistar rats were treated per gavage, during 18 days, with 2.5, 

10 or 90 microg/kg/d astaxanthin dissolved in ethanol (3 g/kg) and CSD was 

recorded on the cortical surface 1 to 3 days thereafter. Four groups, treated 

respectively with ethanol, distilled water and soybean oil with- and without 

astaxanthin were also studied for comparison with the ethanol + astaxanthin 

groups. RESULTS: Ethanol-treated rats displayed higher CSD-velocities (mean 

values, in mm/min, per hour of recording ranging from 4.08 +/- 0.09 to 4.12 +/- 

0.16), compared to the distilled water-group (from 3.19 +/- 0.13 to 3.27 +/- 0.06). 

Addition of astaxanthin to ethanol lead to lower CSD-velocities in a dosedependent 

manner, ranging from 3.68 +/- 0.09 to 3.97 +/- 0.22 for the 2.5 

microg/kg/d-dose, from 3.29 +/- 0.09 to 3.32 +/- 0.07 for the 10 microg/kg/ddose, 

and from 2.89 +/- 0.13 to 2.92 +/- 0.11 for the 90 microg/kg/d-dose. The 

velocities of the soybean oil groups (with and without astaxanthin) were not 

statistically different from the 10 microg/kg/d astaxanthin + ethanol and distilled 

water groups. CONCLUSION: The results demonstrate the antagonistic effect of 

astaxanthin against the ethanol-induced facilitation of CSD propagation. Probably 

carotenoid antioxidant properties are involved in such effects. 




133

Environ Geochem Health. 2007 Dec;29(6):483-9. Epub 2007 Aug 25. 

Impact of astaxanthin-enriched algal powder of Haematococcus 

pluvialis on memory improvement in BALB/c mice. 

Zhang X, Pan L, Wei X, Gao H, Liu J. 

Research and Development Center of Marine Biotechnology, Institute of 

Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao, 266071, 

China. 

The impact of astaxanthin-enriched algal powder on auxiliary memory improvement was 

assessed in BALB/c mice pre-supplemented with different dosages of cracked green algal 

(Haematococcus pluvialis) powder daily for 30 days. The supplemented mice were first 

tested over 8 days to find a hidden platform by swimming in a Morris water maze. Then, 

for 5 days, the mice were used to search for a visible platform in a Morris water maze. 

After that, the mice practised finding a safe place--an insulated platform in a chamber-- 

for 2 days. During these animal experimental periods, similar algal meals containing 

astaxanthin at 0, 0.26, 1.3 and 6.4 mg/kg body weight were continuously fed to each 

group of tested mice. Profiles of latency, distance, speed and the direction angle to the 

platforms as well as the diving frequency in each group were measured and analyzed. 

The process of mice jumping up onto the insulated platform and diving down to the 

copper-shuttered bottom with a 36 V electrical charge were also monitored by automatic 

video recording. The results of the Morris maze experiment showed that middle dosage 

of H. pluvialis meals (1.3 mg astaxanthin/kg body weight) significantly shortened the 

latency and distance required for mice to find a hidden platform. However, there was no 

obvious change in swim velocity in any of the supplemented groups. In contrast, the 

visible platform test showed a significant increase in latency and swim distance, and a 

significant decrease in swim speed for all groups of mice orally supplemented with H. 

pluvialis powder compared to the placebo group (P < 0.05 or P < 0.01). Mice 

supplemented with the algal meal hesitantly turned around the original hidden platform, 

in contract to mice supplemented with placebo, who easily forgot the original location 

and accepted the visible platform as a new safe place. These results illustrate that 

astaxanthin-enriched H. pluvialis powder has the auxiliary property of memory 

improvement. The results from the platform diving test showed that the low and middle 

dosage of H. pluvialis powder, rather that the high dosage, increased the latency and 

reduced the frequency of diving from the safe insulated platform to the electrically 

stimulated copper shutter, especially in the low treatment group (P < 0.05). These results 

indicate that H. pluvialis powder is associated with dose-dependent memory 

improvement and that a low dosage of algal powder (

Effects of astaxanthin on brain damages due to ischemia 

Yoshihisa Kudo': Ryuichi Nakjima+, Nagisa Matsumoto! and Hircki Tsukahara 2 

1. School of Life Science, Tokyo University of Pharmacy and Life Science, 

2. Fujichemical Industry co. ltd 

Brain requires high energy-supply to keep its normal function. Well-developed 

blood vessels in the brain supply enough glucose and oxygen to generate required 

energy. When some part of blood vessels were closed or occluded by some 

reason, the area supported by those blood vessels will fall into ischemia and the 

neuronal cells distributed in the area will be damaged or die. Since neuronal cells 

have no neogenetic properties, the functions supported by the area will be lost 

forever. We know and take care of large scale of neuronal cell death which will 

cause severe loss of brain function, but we ignore small scale of ischemia which 

may have no apparent dysfunction. However senile dementia will be formed due 

to the accumulation of such small scale ischemic neuronal cell death. Although 

big efforts have been made to develop some drugs to rescue the cells exposed to 

ischemia from death, we have no effective drugs so far. Since astaxanthin has 

been known to have antioxidant effects, we expected this drug to rescue the cell 

damage during ischemia and re-perfusion. 

In the present study we used slice preparations (300 urn) of hippocampus obtained 

from young adult rats. To measure intracellular Ca2+ concentration before, during 

and after ischemia we stained the slice preparation by fura-z, a Ca2+ indicator. 

The fluorescence of loaded fura-S was analyzed by an image processor (Argus 

50;Hamamatsu photonics). To examine brain edema during ischemia we used 

self-made devise, which is consisted of an infra-red differential interferance 

microscope with an infra-red camera and an image processor and measured" 

contrast value" as indices ofedema. Astaxanthin (0.003%)pretreated for ten 

minutes before ischemia reduced the increase in intracellular Ca2+ concentration 

duirng ischemia and accelerate the recoveryfrom the abnormal increase in Ca2+ 

concentration Preteated astaxantin (0.01%)alsoreduced the edema 

developedduring ischemia 

Although present results were still preliminary, astaxanthin can be expected 

to have effective rescuing effects on neuronal damages induced by ischemia. 


135

January 2005 Biol. Pharm. Bull. 28(1) 47—52 (2005) 47 

Antihypertensive and Neuroprotective Effects of Astaxanthin in 

Experimental Animals 

Ghazi HUSSEIN,*,a,b Masami NAKAMURA,b Qi ZHAO,b Tomomi IGUCHI,b 

Hirozo GOTO,c Ushio SANKAWA,a and Hiroshi WATANABEb 

a International Research Center for Traditional Medicine, Toyama Prefecture; 

Toyama 939–8224, Japan: b Division of 

Pharmacology; and c Department of Kampo Diagnostics, Institute of Natural 

Medicine, Toyama Medical & 

Pharmaceutical University; Toyama 930–0194, Japan. 

Received May 18, 2004; accepted October 5, 2004 

Astaxanthin is a natural antioxidant carotenoid that occurs in a wide variety of 

living organisms. We investigated, for the first time, antihypertensive effects of 

astaxanthin (ASX-O) in spontaneously hypertensive rats (SHR). Oral 

administration of ASX-O for 14 d induced a significant reduction in the arterial 

blood pressure (BP) in SHR but not in normotensive Wistar Kyoto (WKY) strain. 

The long-term administration of ASX-O (50 mg/kg) for 5 weeks in stroke prone 

SHR (SHR-SP) induced a significant reduction in the BP. It also delayed the 

incidence of stroke in the SHR-SP. To investigate the action mechanism of ASX- 

O, the effects on PGF2a-induced 

contractions of rat aorta treated with NG-nitro-l-arginine methyl ester (L-NAME) 

were studied in vitro. ASX-O (1 to 10mM) induced vasorelaxation mediated by 

nitric oxide (NO). The results suggest that the antihypertensive 

effect of ASX-O may be due to a NO-related mechanism. ASX-O also showed 

significant neuroprotective effects in ischemic mice, presumably due to its 

antioxidant potential. Pretreatment of the mice with ASX-O significantly 

shortened the latency of escaping onto the platform in the Morris water maze 

learning performance test. In conclusion, these results indicate that astaxanthin 

can exert beneficial effects in protection against hypertension and stroke and in 

improving memory in vascular dementia. 


136

Food Chem Toxicol. 2010 Jun;48(6):1741-5. Epub 2010 Apr 9. 

Astaxanthin improves the proliferative capacity as well as the osteogenic 

and adipogenic differentiation potential in neural stem cells. 

Kim JH, Nam SW, Kim BW, Kim WJ, Choi YH. 

Department of Biomaterial Control, Dong-Eui University, Busan, South Korea. 


In the present study, the effect of astaxanthin on improvement of the proliferative 

capacity as well as the osteogenic and adipogenic differentiation potential in neural stem 

cells (NSCs) was evaluated. Treatment of astaxanthin-induced actives cell growth in a 

dose-dependent and time-dependent manner. Results from a clonogenic assay clearly 

indicated that astaxanthin can actively stimulate proliferation of NSCs. Astaxanthininduced 

improvement in the proliferative capacity of NSCs resulted in overexpression of 

several proliferation-related proteins. Astaxanthin-induced activation of PI3K and its 

downstream mediators, p-MEK, p-ERK, and p-Stat3 in NSCs resulted in subsequent 

induction of expression of proliferation-related transcription factors (Rex1, CDK1, and 

CDK2) and stemness genes (OCT4, SOX2, Nanog, and KLF4). Astaxanthin also 

improved the osteogenic and adipogenic differentiation potential of NSCs. Astaxanthintreated 

NSCs showed prominent calcium deposits and fat formation. These results were 

consistent with overexpression of osteogenesis-related genes (osteonectin, RXR, and 

osteopontin) and adipogenesis-related genes (AP and PPAR-gamma) after astaxanthin 

treatment. These findings clearly demonstrated that astaxanthin acts synergistically on the 

regulatory circuitry that controls proliferation and differentiation of NSCs. Copyright 

2010 Elsevier Ltd. All rights reserved. 



137

Brain Res. 2010 Sep 7. [Epub ahead of print] 

Astaxanthin upregulates heme oxygenase-1 expression through ERK1/2 

pathway and its protective effect against beta-amyloid-induced 

cytotoxicity in SH-SY5Y cells. 

Wang HQ, Sun XB, Xu YX, Zhao H, Zhu QY, Zhu CQ. 


Astaxanthin (ATX), the most abundant flavonoids in propolis, has been proven to exert 

neuroprotective property against glutamate-induced neurotoxicity and ischemiareperfusion-induced 

apoptosis. Previous study have revealed that ATX can rescue PC12 

cells from Aβ(25-35)-induced apoptotic death. However, the mechanisms by which ATX 

mediates its therapeutic effects in vitro are unclear. In the present study, we explored the 

underlying mechanisms involved in the protective effects of ATX on the Aβ(25-35)- 

induced cytotoxicity in SH-SY5Y cells. Pre-treatment with ATX for 4h significantly 

reduced the Aβ(25-35)-induced viability loss, apoptotic rate and attenuated Aβ-mediated 

ROS production. In addition, ATX inhibited Aβ(25-35)-induced lowered membrane 

potential, decreased Bcl-2/Bax ratio. We also demonstrated that ATX could prevent the 

activation of p38MAPK kinase pathways induced by Aβ. Moreover, we for the first time 

have revealed the ATX increased antioxidant enzyme heme oxygenase-1 (HO-1) 

expression in concentration-dependent and time-dependent manners, which were 

correlated with its protective effect against Aβ(25-35)-induced injury. Because the 

inhibitor of HO-1 activity, ZnPP reversed the protective effect of ATX against Aβ(25- 

35)-induced cell death. We also demonstrated that the specific ERK inhibitor, PD98059, 

concentration-dependently blocked on ATX-induced HO-1 expression, and meanwhile 

PD98059 reversed the protective effect of ATX against Aβ25-35-induced cell death. 

Taken together, these findings suggest that astaxanthin can induce HO-1 expression 

through activation of ERK signal pathways, thereby protecting the SH-SY5Y cells from 

Aβ(25-35)-induced oxidative cell death. 



138

J Clin Biochem Nutr. 2009 May;44(3):280-4. Epub 2009 Apr 25. 

Preliminary Clinical Evaluation of Toxicity and Efficacy of A New 

Astaxanthin-rich Haematococcus pluvialis Extract. 

Satoh A, Tsuji S, Okada Y, Murakami N, Urami M, Nakagawa K, Ishikura M, Katagiri 

M, Koga Y, Shirasawa T. 


Astaxanthin (Ax), a carotenoid ubiquitously distributed in microorganisms, fish, and 

crustaceans, has been known to be a potent antioxidant and hence exhibit various 

physiological effects. We attempted in these studies to evaluate clinical toxicity and 

efficacy of long-term administration of a new Ax product, by measuring biochemical and 

hematological blood parameters and by analyzing brain function (using CogHealth and 

P300 measures). Ax-rich Haematococcus pluvialis extracts equivalent to 4, 8, 20 mg of 

Ax dialcohol were administered to 73, 38, and 16 healthy adult volunteers, respectively, 

once daily for 4 weeks to evaluate safety. Ten subjects with age-related forgetfulness 

received an extract equivalent to 12 mg in a daily dosing regimen for 12 weeks to 

evaluate efficacy. As a result, no abnormality was observed and efficacy for age-related 

decline in cognitive and psychomotor functions was suggested. 

PMID: 19430618 [PubMed - in process]PMCID: PMC2675019Free PMC Article 


139

J Food Sci. 2009 Sep;74(7):H225-31. 

Antioxidative and anti-inflammatory neuroprotective effects of 

astaxanthin and canthaxanthin in nerve growth factor differentiated 

PC12 cells. 

Chan KC, Mong MC, Yin MC. 

Dept of Food and Nutrition, Providence Univ, Taichung County, Taiwan. 


Nerve growth factor differentiated PC12 cells were used to examine the antioxidative and 

anti-inflammatory effects of astaxanthin (AX) and canthaxanthin (CX). PC12 cells were 

pretreated with AX or CX at 10 or 20 muM, and followed by exposure of hydrogen 

peroxide (H(2)O(2)) or 1-methyl-4-phenylpyridinium ion (MPP(+)) to induce cell injury. 

H(2)O(2) or MPP(+) treatment significantly decreased cell viability, increased lactate 

dehydrogenase (LDH) release, enhanced DNA fragmentation, and lowered mitochondrial 

membrane potential (MMP) (P < 0.05). The pretreatments from AX or CX concentrationdependently 

alleviated H(2)O(2) or MPP(+)-induced cell death, LDH release, DNA 

fragmentation, and MMP reduction (P < 0.05). Either H(2)O(2) or MPP(+) treatment 

significantly increased malonyldialdehyde (MDA) and reactive oxygen species (ROS) 

formations, decreased glutathione content, and lowered glutathione peroxidase (GPX) 

and catalase activities (P < 0.05). The pretreatments from AX or CX significantly 

retained GPX and catalase activities, and decreased MDA and ROS formations (P < 

0.05). H(2)O(2) or MPP(+) treatment significantly decreased Na(+)-K(+)-ATPase 

activity, elevated caspase-3 activity and levels of interleukin (IL)-1, IL-6, and tumor 

necrosis factor (TNF)-alpha (P < 0.05); and the pretreatments from these agents 

significantly restored Na(+)-K(+)-ATPase activity, suppressed caspase-3 activity and 

release of IL-1, IL-6, and TNF-alpha (P < 0.05). Based on the observed antioxidative and 

anti-inflammatory protection from AX and CX, these 2 compounds were potent agents 

against neurodegenerative disorder. 



140

J Clin Biochem Nutr. 2010 Sep;47(2):121-9. Epub 2010 Jul 6. 

Neuroprotective Effects of Astaxanthin in Oxygen-Glucose Deprivation 

in SH-SY5Y Cells and Global Cerebral Ischemia in Rat. 

Lee DH, Lee YJ, Kwon KH. 

Departments of Surgery and Pharmacology, School of Medicine, University of 

Pittsburgh, Pittsburgh, Pennsylvania 15213, USA. 


Astaxanthin (ATX), a naturally occurring carotenoid pigment, is a powerful biological 

antioxidant. In the present study, we investigated whether ATX pharmacologically offers 

neuroprotection against oxidative stress by cerebral ischemia. We found that the 

neuroprotective efficacy of ATX at the dose of 30 mg/kg (n = 8) was 59.5% compared 

with the control group (n = 3). In order to make clear the mechanism of ATX 

neuroprotection, the up-regulation inducible nitric oxide synthase (iNOS) and heat shock 

proteins (HSPs) together with the oxygen glucose deprivation (OGD) in SH-SY5Y cells 

were also investigated. The induction of various factors involved in oxidative stress 

processes such as iNOS was suppressed by the treatment of ATX at 25 and 50 µM after 

OGD-induced oxidative stress. In addition, Western blots showed that ATX elevated of 

heme oxygenase-1 (HO-1; Hsp32) and Hsp70 protein levels in in vitro. These results 

suggest that the neuroprotective effects of ATX were related to anti-oxidant activities in 

global ischemia. 

PMID: 20838567 [PubMed - in process]PMCID: PMC2935152 


141

J Med Food. 2010 Jun;13(3):548-56. 

Astaxanthine secured apoptotic death of PC12 cells induced by betaamyloid 

peptide 25-35: its molecular action targets. 

Chang CH, Chen CY, Chiou JY, Peng RY, Peng CH. 

Research Institute of Biotechnology, Hungkuang University, Taichung Hsien, Taiwan. 


Astaxanthine (ASTx) is a novel carotenoid nutraceutical occurring in many crustaceans 

and red yeasts. It has potent antioxidant, photoprotective, hepatodetoxicant, and antiinflammatory 

activities. Documented effect of ASTx on treatment of neurodegenerative 

disease is still lacking. We used the beta-amyloid peptide (Abeta) 25-35-treated PC12 

model to investigate the neuron-protective effect of ASTx. The parameters examined 

included cell viability, caspase activation, and various apoptotic biomarkers that play 

their critical roles in the transduction pathways independently or synergistically. Results 

indicated that Abeta25-35 at 30 microM suppressed cell viability by 55%, whereas ASTx 

was totally nontoxic below a dose of 5.00 microM. ASTx at 0.1 microM protected PC12 

cells from damaging effects of Abeta25-35 in several ways: (1) by securing the cell 

viability; (2) by partially down-regulating the activation of caspase 3; (3) by inhibiting 

the expression of Bax; (4) by completely eliminating the elevation of interleukin-1beta 

and tumor necrosis factor-alpha; (5) by inhibiting the nuclear translocation of nuclear 

factor kappaB; (6) by completely suppressing the phosphorylation of p38 mitogenactivated 

protein kinase; (7) by completely abolishing the calcium ion influx to 

effectively maintain calcium homeostasis; and (8) by suppressing the majority (about 

75%) of reactive oxygen species production. Conclusively, ASTx may have merit to be 

used as a very potential neuron protectant and an anti-early-stage Alzheimer's disease 

adjuvant therapy. 



142

Brain Res. 2010 Sep 21. [Epub ahead of print] 

Neuroprotective effect of astaxanthin on H(2)O(2)-induced 

neurotoxicity in vitro and on focal cerebral ischemia in vivo. 

Lu YP, Liu SY, Sun H, Wu XM, Li JJ, Zhu L. 

Institute of Nautical Medicine, Nantong University, Nantong 226001, China. 


Astaxanthin (AST) is a powerful antioxidant that occurs naturally in a wide variety of 

living organisms. Much experimental evidence has proved that AST has the function of 

eliminating oxygen free radicals and can protect organisms from oxidative damage. The 

present study was carried out to further investigate the neuroprotective effect of AST on 

oxidative stress induced toxicity in primary culture of cortical neurons and on focal 

cerebral ischemia-reperfusion induced brain damage in rats. AST, over a concentration 

range of 250-1000nM, attenuated 50μM H(2)O(2)-induced cell viability loss. 500nM 

AST pretreatment significantly inhibited H(2)O(2)-induced apoptosis measured by 

Hoechst 33342 staining and restored the mitochondrial membrane potential (MMP) 

measured by a fluorescent dye, Rhodamine 123. In vivo, AST prevented cerebral 

ischemic injury induced by 2h middle cerebral artery occlusion (MCAO) and 24h 

reperfusion in rats. Pretreatment of AST intragastrically twice at 5h and 1h prior to 

ischemia dramatically diminished infarct volume and improved neurological deficit in a 

dose-dependent manner. Nissl staining showed that the neuronal injury was significantly 

improved by pretreatment of AST at 80mg/kg. Taken together, these results suggest that 

pretreatment with AST exhibits noticeable neuroprotection against brain damage induced 

by ischemia-reperfusion and the antioxidant activity of AST maybe partly responsible for 

it. 



143

J Microbiol Biotechnol. 2009 Nov;19(11):1355-63. 

Astaxanthin inhibits H2O2-mediated 

apoptotic cell death in mouse neural 

progenitor cells via modulation of P38 

and MEK signaling pathways. 

Kim JH, Choi W, Lee JH, Jeon SJ, Choi YH, Kim BW, Chang HI, Nam SW. 

Source 

Department of Biomaterial Control,Dong-Eui University, Busan 614-714, Korea. 


In the present study, neuroprotective effects of astaxanthin on H2O2-mediated apoptotic 

cell death using cultured mouse neural progenitor cells (mNPCs) were investigated. To 

cause apoptotic cell death, mNPCs were pretreated with astaxanthin for 8 h and followed 

by treatment of 0.3 mM H2O2. Pretreatment of mNPCs with astaxanthin significantly 

inhibited H2O2-mediated apoptosis and induced cell growth in a dose-dependent manner. 

In Western blot analysis, astaxanthin-pretreated cells showed the activation of p-Akt, p- 

MEK, p-ERK, and Bcl-2, and the reduction of p-P38, p-SAPK/JNK, Bax, p-GSK3beta, 

cytochrome c, caspase-3, and PARP. Because H2O2 triggers caspases activation, this 

study examined whether astaxanthin can inhibit caspases activation in H2O2-treated 

mNPCs. After H2O2 treatment, caspases activities were prominently increased but 

astaxanthin pretreatment significantly inhibited H2O2-mediated caspases activation. 

Astaxanthin pretreatment also significantly recovered ATP production ability of H2O2- 

treated cells. These findings indicate that astaxanthin inhibits H2O2-mediated apoptotic 

features in mNPCs. Inhibition assays with SB203580 (10 microM, a specific inhibitor of 

p38) and PD98059 (10 microM, a specific inhibitor of MEK) clearly showed that 

astaxanthin can inhibit H2O2-mediated apoptotic death via modulation of p38 and MEK 

signaling pathways. 



144

Pharmacol Biochem Behav. 2011 Sep;99(3):349-55. Epub 2011 May 17. 

Astaxanthin limits fish oil-related 

oxidative insult in the anterior forebrain 

of Wistar rats: putative anxiolytic effects? 

Mattei R, Polotow TG, Vardaris CV, Guerra BA, Leite JR, Otton R, Barros MP. 

Source 

Department of Psychobiology, Universidade Federal de São Paulo (UNIFESP), ZIP 

04023062, São Paulo, SP, Brazil. 


The habitual consumption of marine fish is largely associated to human mental health. 

Fish oil is particularly rich in n-3 polyunsaturated fatty acids that are known to play a role 

in several neuronal and cognitive functions. In parallel, the orange-pinkish carotenoid 

astaxanthin (ASTA) is found in salmon and displays important antioxidant and antiinflammatory 

properties. Many neuronal dysfunctions and anomalous psychotic behavior 

(such as anxiety, depression, etc.) have been strongly related to the higher sensitivity of 

cathecolaminergic brain regions to oxidative stress. Thus, the aim of this work was to 

study the combined effect of ASTA and fish oil on the redox status in plasma and in the 

monoaminergic-rich anterior forebrain region of Wistar rats with possible correlations 

with the anxiolytic behavior. Upon fish oil supplementation, the downregulation of 

superoxide dismutase and catalase activities combined to increased "free" iron content 

resulted in higher levels of lipid and protein oxidation in the anterior forebrain of 

animals. Such harmful oxidative modifications were hindered by concomitant 

supplementation with ASTA despite ASTA-related antioxidant protection was mainly 

observed in plasma. Although it is clear that ASTA properly crosses the brain-blood 

barrier, our data also address a possible indirect role of ASTA in restoring basal oxidative 

conditions in anterior forebrain of animals: by improving GSH-based antioxidant 

capacity of plasma. Preliminary anxiolytic tests performed in the elevated plus maze are 

in alignment with our biochemical observations. 

Copyright © 2011 Elsevier Inc. All rights reserved. 



145

Biofactors. 2011 Jan;37(1):25-30. doi: 10.1002/biof.130. Epub 2010 Nov 11. 

The antianxiety-like effect of astaxanthin 

extracted from Paracoccus 

carotinifaciens. 

Nishioka Y, Oyagi A, Tsuruma K, Shimazawa M, Ishibashi T, Hara H. 

Source 

Molecular Pharmacology, Department of Biofunctional Evaluation, Gifu Pharmaceutical 

University, 1-25-4 Daigaku-nishi, Gifu, Japan. 


Astaxanthin is a red carotenoid pigment and is widely found in living organisms. 

Astaxanthin has a potent antioxidative ability and has been reported as having various 

biological effects on the central nerve system, such as a protective effect against 

ischemia/reperfusion injury and improvement in cognitive function. In this study, to 

investigate the effects of astaxanthin on anxiety and depression, we performed some 

behavioral trials including the elevated plus maze test, hole-board test, forced swim test, 

and tail suspension test. Astaxanthin (100 and 300 mg/kg/day for 10 days, p.o.) 

significantly increased the time spent in open arms in the elevated plus maze test and 

increased the head-dipping count and duration in the hole-board test. On the other hand, 

astaxanthin (10, 100, 300, and 500 mg/kg/day for 10 days, p.o.) did not change the 

immobility time in the forced swim test or the tail suspension test. In conclusion, in mice, 

astaxanthin exerted anxiolytic-like effects, but not antidepressant-like effects. 

Copyright © 2010 International Union of Biochemistry and Molecular Biology, Inc. 



146

Food Chem Toxicol. 2011 Jan;49(1):271-80. Epub 2010 Nov 5. 

Astaxanthin protects against MPTP/MPP+-induced 

mitochondrial dysfunction and ROS production in vivo 

and in vitro. 

Lee DH, Kim CS, Lee YJ. 

Source 

Department of Surgery, School of Medicine, University of Pittsburgh, Pittsburgh, PA 

15213, USA. 


Astaxanthin (AST) is a powerful antioxidant that occurs naturally in a wide variety of 

living organisms. We have investigated the role of AST in preventing 1-methyl-4-phenyl- 

1,2,3,6-tetrahydropyridine (MPTP)-induced apoptosis of the substantia nigra (SN) 

neurons in the mouse model of Parkinson's disease (PD) and 1-methyl-4- 

phenylpyridinium (MPP+)-induced cytotoxicity of SH-SY5Y human neuroblastoma 

cells. In in vitro study, AST inhibits MPP+-induced production of intracellular reactive 

oxygen species (ROS) and cytotoxicity in SH-SY5Y human neuroblastoma cells. 

Preincubation of AST (50 μM) significantly attenuates MPP+-induced oxidative damage. 

Furthermore, AST is able to enhance the expression of Bcl-2 protein but reduce the 

expression of α-synuclein and Bax, and suppress the cleavage of caspase-3. Our results 

suggest that the protective effects of AST on MPP+-induced apoptosis may be due to its 

anti-oxidative properties and anti-apoptotic activity via induction of expression of 

superoxide dismutase (SOD) and catalase and regulating the expression of Bcl-2 and 

Bax. Pretreatment with AST (30 mg/kg) markedly increases tyrosine hydroxylase (TH)- 

positive neurons and decreases the argyrophilic neurons compared with the MPTP model 

group. In summary, AST shows protection from MPP+/MPTP-induced apoptosis in the 

SH-SY5Y cells and PD model mouse SN neurons, and this effect may be attributable to 

upregulation of the expression of Bcl-2 protein, downregulation of the expression of Bax 

and α-synuclein, and inhibition of the activation of caspase-3. These data indicate that 

AST may provide a valuable therapeutic strategy for the treatment of progressive 

neurodegenerative disease such as Parkinson's disease. 

Copyright © 2010 Elsevier Ltd. All rights reserved. 


PMCID: PMC3010303 [Available on 2012/1/1] 


147

Int J Mol Sci. 2010;11(12):5109-19. Epub 2010 Dec 9. 

Astaxanthin Improves Stem Cell Potency 

via an Increase in the Proliferation of 

Neural Progenitor Cells. 

Kim JH, Nam SW, Kim BW, Choi W, Lee JH, Kim WJ, Choi YH. 

Source 

Department of Biomaterial Control, Dong-Eui University, Busan, 614-714, Korea; E- 

Mails: 12845@deu.ac.kr (J.-H.K.); bwkim@deu.ac.kr (B.-W.K.); wbchoi@deu.ac.kr 

(W.C.); jonghwanlee@deu.ac.kr (J.-H.L.). 


The present study was designed to investigate the question of whether or not astaxanthin 

improves stem cell potency via an increase in proliferation of neural progenitor cells 

(NPCs). Treatment with astaxanthin significantly increased proliferation and colony 

formation of NPCs. For identification of possible activated signaling molecules involved 

in active cell proliferation occurring after astaxanthin treatment, total protein levels of 

several proliferation-related proteins, and expression levels of proliferation-related 

transcription factors, were assessed in NPCs. In Western blot analysis, astaxanthin 

induced significant activation of phosphatidylinositol 3-kinase (PI3K) and its 

downstream mediators in a time-dependent manner. Results of RT-PCR analysis showed 

upregulation of proliferation-related transcription factors and stemness genes. To estimate 

the relevance of PI3K and mitogen-activated protein, or extracellular signal-regulated 

kinase kinase (MEK) signaling pathways in cell growth of astaxanthin-treated NPCs, 

inhibition assays were performed with LY294002, a specific inhibitor of PI3K, and 

PD98059, a specific inhibitor of MEK, respectively. These results clearly showed that 

astaxanthin induces proliferation of NPCs via activation of the PI3K and MEK signaling 

pathways and improves stem cell potency via stemness acting signals. 




148

Cardioprotective 

Atherosclerosis. 2010 Apr;209(2):520-3. Epub 2009 Oct 14. 

Administration of natural astaxanthin increases serum HDL-cholesterol 

and adiponectin in subjects with mild hyperlipidemia. 

Yoshida H, Yanai H, Ito K, Tomono Y, Koikeda T, Tsukahara H, Tada N. 

Department of Laboratory Medicine, Jikei University Kashiwa Hospital, Chiba, Japan. 

hyoshida@jikei.ac.jp 


BACKGROUND: Astaxanthin has been reported to improve dyslipidemia and metabolic 

syndrome in animals, but such effects in humans are not well known. 

METHODS: Placebo-controlled astaxanthin administration at doses of 0, 6, 12, 18 

mg/day for 12 weeks was randomly allocated to 61 non-obese subjects with fasting serum 

triglyceride of 120-200mg/dl and without diabetes and hypertension, aged 25-60 years. 

RESULTS: In before and after tests, body mass index (BMI) and LDL-cholesterol were 

unaffected at all doses, however, triglyceride decreased, while HDL-cholesterol increased 

significantly. Multiple comparison tests showed that 12 and 18 mg/day doses 

significantly reduced triglyceride, and 6 and 12 mg doses significantly increased HDLcholesterol. 

Serum adiponectin was increased by astaxanthin (12 and 18 mg/day), and 

changes of adiponectin correlated positively with HDL-cholesterol changes independent 

of age and BMI. 

CONCLUSIONS: This first-ever randomized, placebo-controlled human study suggests 

that astaxanthin consumption ameliorates triglyceride and HDL-cholesterol in correlation 

with increased adiponectin in humans. 



149

Am J Cardiol. 2008 May 22;101(10A):58D-68D. 

Astaxanthin: a novel potential treatment for oxidative stress and 

inflammation in cardiovascular disease. 

Pashkow FJ, Watumull DG, Campbell CL. 

John A. Burns School of Medicine, University of Hawaii, Honolulu, Hawaii, 

USA. fpashkow@cardaxpharma.com 

Oxidative stress and inflammation are implicated in several different 

manifestations of cardiovascular disease (CVD). They are generated, in part, from 

the overproduction of reactive oxygen species (ROS) and reactive nitrogen 

species (RNS) that activate transcriptional messengers, such as nuclear factorkappaB, 

tangibly contributing to endothelial dysfunction, the initiation and 

progression of atherosclerosis, irreversible damage after ischemic reperfusion, and 

even arrhythmia, such as atrial fibrillation. Despite this connection between 

oxidative stress and CVD, there are currently no recognized therapeutic 

interventions to address this important unmet need. Antioxidants that provide a 

broad, "upstream" approach via ROS/RNS quenching or free radical chain 

breaking seem an appropriate therapeutic option based on epidemiologic, dietary, 

and in vivo animal model data. However, human clinical trials with several 

different well-known agents, such as vitamin E and beta-carotene, have been 

disappointing. Does this mean antioxidants as a class are ineffective, or rather that 

the "right" compound(s) have yet to be found, their mechanisms of action 

understood, and their appropriate targeting and dosages determined? A large class 

of potent naturally-occurring antioxidants exploited by nature-the oxygenated 

carotenoids (xanthophylls)-have demonstrated utility in their natural form but 

have eluded development as successful targeted therapeutic agents up to the 

present time. This article characterizes the mechanism by which this novel group 

of antioxidants function and reviews their preclinical development. Results from 

multiple species support the antioxidant/anti-inflammatory properties of the 

prototype compound, astaxanthin, establishing it as an appropriate candidate for 

development as a therapeutic agent for cardiovascular oxidative stress and 

inflammation. 


 

Review 



150

Am J Cardiol. 2008 May 22;101(10A):20D-29D. 

Biologic activity of carotenoids related to distinct membrane 

physicochemical interactions. 

McNulty H, Jacob RF, Mason RP. 

Elucida Research, Beverly, MA 01915, USA. 

Carotenoids are naturally occurring organic pigments that are believed to have 

therapeutic benefit in treating cardiovascular disease (CVD) because of their 

antioxidant properties. However, prospective randomized trials have failed to 

demonstrate a consistent benefit for the carotenoid beta-carotene in patients at risk 

for CVD. The basis for this apparent paradox is not well understood but may be 

attributed to the distinct antioxidant properties of various carotenoids resulting 

from their structure-dependent physicochemical interactions with biologic 

membranes. To test this hypothesis, we measured the effects of astaxanthin, 

zeaxanthin, lutein, beta-carotene, and lycopene on lipid peroxidation using model 

membranes enriched with polyunsaturated fatty acids. The correlative effects of 

these compounds on membrane structure were determined using small-angle x- 

ray diffraction approaches. The nonpolar carotenoids, lycopene and beta-carotene, 

disordered the membrane bilayer and stimulated membrane lipid peroxidation 

(>85% increase in lipid hydroperoxide levels), whereas astaxanthin (a polar 

carotenoid) preserved membrane structure and exhibited significant antioxidant 

activity (>40% decrease in lipid hydroperoxide levels). These results suggest that 

the antioxidant potential of carotenoids is dependent on their distinct membrane 

lipid interactions. This relation of structure and function may explain the 

differences in biologic activity reported for various carotenoids, with important 

therapeutic implications. 




151

Mol Cell Biochem. 2008 Feb;309(1-2):61-8. Epub 2007 Nov 16. 

The protective role of carotenoids against 7-keto-cholesterol 

formation in solution. 

Palozza P, Barone E, Mancuso C, Picci N. 

Institute of General Pathology, Catholic University School of Medicine, Largo F. 

Vito 1, 00168 Rome, Italy. p.palozza@rm.unicatt.it 

The antioxidant activity of beta-carotene and oxygenated carotenoids lutein, 

canthaxanthin, and astaxanthin was investigated during spontaneous and peroxylradical-induced 

cholesterol oxidation. Cholesterol oxidation, measured as 

generation of 7-keto-cholesterol (7-KC), was evaluated in a heterogeneous 

solution with cholesterol, AAPH, and carotenoids solubilized in tetrahydrofuran 

and in water, and in a homogeneous solution of chlorobenzene, with AIBN as a 

prooxidant. The formation of 7-KC was dependent on temperature and on 

cholesterol and prooxidant concentrations. All the carotenoids tested, exhibited 

significant antioxidant activity by inhibiting spontaneous, AAPH- and AIBNinduced 

formation of 7-KC, although the overall order of efficacy of these 

compounds was astaxanthin > canthaxanthin > lutein = beta-carotene. The finding 

that carotenoids exert protective effects on spontaneous and free radical-induced 

cholesterol oxidation may have important beneficial effects on human health, by 

limiting the formation of atheroma and by inhibiting cholesterol oxidation in food 

processing or storage. 




152

Int J Vitam Nutr Res. 2007 Jan;77(1):3-11. 

Effects of astaxanthin supplementation on lipid peroxidation. 

Karppi J, Rissanen TH, Nyyssönen K, Kaikkonen J, Olsson AG, Voutilainen 

S, Salonen JT. 

Research Institute of Public Health, University of Kuopio, P.O. Box 1627, FI- 

70211 Kuopio, Finland. jouni.karppi@uku.fi 

Astaxanthin, the main carotenoid pigment in aquatic animals, has greater 

antioxidant activity in vitro (protecting against lipid peroxidation) and a more 

polar configuration than other carotenoids.We investigated the effect of threemonth 

astaxanthin supplementation on lipid peroxidation in healthy non-smoking 

Finnish men, aged 19-33 years by using a randomized double-blind study design. 

Also absorption of astaxanthin from capsules into bloodstream and its safety were 

evaluated. The intervention group received two 4-mg astaxanthin (Astaxin) 

capsules daily, and the control group two identical-looking placebo capsules. 

Astaxanthin supplementation elevated plasma astaxanthin levels to 0.032 pmol/L 

(p < 0.001 for the change compared with the placebo group). We observed that 

levels of plasma 12- and 15-hydroxy fatty acids were reduced statistically 

significantly in the astaxanthin group (p = 0.048 and p = 0.047 respectively) 

during supplementation, but not in the placebo group and the change of 15- 

hydroxy fatty acid was almost significantly greater (p = 0.056) in the astaxanthin 

group, as compared with the placebo group. The present study suggests that 

intestinal absorption of astaxanthin delivered as capsules is adequate, and well 

tolerated. Supplementation with astaxanthin may decrease in vivo oxidation of 

fatty acids in healthy men. 




153

BMC Nephrol. 2008 Dec 18;9:17. 

Astaxanthin vs placebo on arterial stiffness, oxidative stress and 

inflammation in renal transplant patients (Xanthin): a randomised 

controlled trial. 

Fassett RG, Healy H, Driver R, Robertson IK, Geraghty DP, Sharman JE, 

Coombes JS. 

Renal Research, Royal Brisbane and Women's Hospital, Herston, Brisbane, 

Queensland, Australia. rfassett@mac.com 

BACKGROUND: There is evidence that renal transplant recipients have 

accelerated atherosclerosis manifest by increased cardiovascular morbidity and 

mortality. The high incidence of atherosclerosis is, in part, related to increased 

arterial stiffness, vascular dysfunction, elevated oxidative stress and inflammation 

associated with immunosuppressive therapy. The dietary supplement astaxanthin 

has shown promise as an antioxidant and anti-inflammatory therapeutic agent in 

cardiovascular disease. The aim of this trial is to investigate the effects of 

astaxanthin supplementation on arterial stiffness, oxidative stress and 

inflammation in renal transplant patients. METHOD AND DESIGN: This is a 

randomised, placebo controlled clinical trial. A total of 66 renal transplant 

recipients will be enrolled and allocated to receive either 12 mg/day of 

astaxanthin or an identical placebo for one-year. Patients will be stratified into 

four groups according to the type of immunosuppressant therapy they receive: 1) 

cyclosporine, 2) sirolimus, 3) tacrolimus or 4) prednisolone+/-azathioprine, 

mycophenolate mofetil or mycophenolate sodium. Primary outcome measures 

will be changes in 1) arterial stiffness measured by aortic pulse wave velocity 

(PWV), 2) oxidative stress assessed by plasma isoprostanes and 3) inflammation 

by plasma pentraxin 3. Secondary outcomes will include changes in vascular 

function assessed using the brachial artery reactivity (BAR) technique, carotid 

artery intimal medial thickness (CIMT), augmentation index (AIx), left 

ventricular afterload and additional measures of oxidative stress and 

inflammation. Patients will undergo these measures at baseline, six and 12 

months. DISCUSSION: The results of this study will help determine the efficacy 

of astaxanthin on vascular structure, oxidative stress and inflammation in renal 

transplant patients. This may lead to a larger intervention trial assessing 

cardiovascular morbidity and mortality. TRIAL REGISTRATION: 

ACTRN12608000159358. 




154

J Clin Biochem Nutr. 2008 Sep;43(2):69-74. 

Effects of astaxanthin on human blood rheology. 

Miyawaki H, Takahashi J, Tsukahara H, Takehara I. 

Miyawaki Orthopedic Clinic, Hokkaido 061-1431, Japan. 

Effects of astaxanthin (AX) derived from H. pluvialis on human blood rheology 

were investigated in 20 adult men with a single-blind method. The experimental 

group was 57.5 +/- 9.8 years of age and the placebo group was 50.8 +/- 13.1 years 

of age. A blood rheology test that measures whole blood transit time was 

conducted using heparinized blood of the volunteers by a MC-FAN apparatus 

(microchannel array flow analyzer). After administration of AX 6 mg/day for 10 

days, the values of the experimental group were decreased from 52.8 +/- 4.9 s to 

47.6 +/- 4.2 s (p

Pharmacology. 2008;82(1):67-73. Epub 2008 May 14. 

Disodium disuccinate astaxanthin prevents carotid artery 

rethrombosis and ex vivo platelet activation. 

Lauver DA, Driscoll EM, Lucchesi BR. 

Department of Pharmacology, University of Michigan Medical School, Ann 

Arbor, Mich 48109, USA. 

BACKGROUND/AIMS:The disodium disuccinate derivative of astaxanthin 

(DDA) is a carotenoid antioxidant under development for the treatment of 

ischemic cardiovascular events. Recent evidence suggests that reactive oxygen 

species (ROS) play an important role in platelet activation. This study seeks to 

investigate the effects of a reactive oxygen species quencher, DDA, in a canine 

model of carotid artery thrombosis. METHODS: After formation of an occlusive 

carotid thrombus, dogs were administered recombinant tissue plasminogen 

activator intra-arterially to achieve thrombolysis in the presence of either 0.9% 

NaCl solution or DDA (10-50 mg/kg i.v. infusion). Ex vivo platelet aggregation 

and tongue bleeding times were measured before and after drug administration. 

Residual thrombus mass was analyzed at the end of each experiment. 

RESULTS:The data indicated a dose- dependent reduction in the incidence of 

carotid artery rethrombosis. In addition, platelet aggregation and thrombus 

weights were dose-dependently inhibited by DDA. No change was recorded in 

tongue bleeding time among the treatment groups. CONCLUSIONS:The data 

demonstrate that at the doses used in this study, DDA significantly reduced the 

incidence of secondary thrombosis while maintaining normal hemostasis. The 

results suggest that upon further study, DDA may one day find utility in 

revascularization procedures. Copyright 2008 S. Karger AG, Basel. 



156

Arzneimittelforschung. 2007;57(1):26-30. 

Eulipidemic effects of berberine administered alone or in 

combination with other natural cholesterol-lowering agents. A 

single-blind clinical investigation. 

Cicero AF, Rovati LC, Setnikar I. 

"G. Descovich" Atherosclerosis and Dysmetabolic Disease Research Center, "D. 

Campanacci" Clinical Medicine and Applied Biotechnology Department, 

University of Bologna, Bologna, Italy. 

Berberine (BERB) and a combination (COMB) of berberine (CAS 2086-83-1) 

with policosanol (CAS 557-61-9), red yeast extract (containing monacolin, CAS 

557-61-9), folic acid and astaxanthin were orally administered daily for 4 weeks 

to 40 subjects with moderate dyslipidemias divided in two parallel groups each of 

20 subjects. Total cholesterol (TC), LDL, HDL, Non HDL, ApoB, ApoA, Lp(a) 

and triglycerides (TG) were measured before and at the end of treatments. BERB 

and COMB significantly reduced TC (respectively by 16% and 20%), LDL (by 

20% and 25%), ApoB (by 15% and 29%) and TG (by 22% and 26%), and 

increased HDL (by 6.6% and 5.1%). Adverse events or impairments of liver 

transaminases or of CPK were not observed. In conclusion, food supplements 

containing natural products such as berberine, policosanol, red yeast extracts, 

folic acid and astaxanthin could be a useful support to diet and life style changes 

to correct dyslipidemias and to reduce cardiovascular risk in subjects with 

moderate mixed dyslipidemias. 




157

Cardiovasc Hematol Agents Med Chem. 2006 Oct;4(4):335-49. 

Retrometabolic syntheses of astaxanthin (3,3'-dihydroxy-beta,beta-carotene-4,4'- 

dione) conjugates: a novel approach to oral and parenteral cardio-protection. 

Lockwood SF, Jackson HL, Gross GJ. 

Hawaii Biotech, Inc., 99-193 Aiea Heights Drive, Suite 200, Aiea, HI 96701, USA. 


Disodium disuccinate astaxanthin has potent cardioprotective effects in animals, 

with demonstrated preclinical efficacy in the rat, rabbit, and canine models of 

experimental infarction. It has been effective in subchronic and acute dosing 

regimens after parenteral administration, and recently published data in rats 

demonstrate that oral cardioprotection is also readily achieved. Myocardial 

salvage in the canine can reach 100% with a 4-day subchronic dosing regimen; 

single-dose I.V. cardioprotection, when given 2 hours before experimental 

coronary occlusion, is on average two-thirds of that achieved with the subchronic 

regimen in dogs. In conscious animals, no effects on hemodynamic parameters 

have been observed. Recently, the beneficial properties of this prototypical 

astaxanthin conjugate have been extended to include second- and third-generation 

compounds with improved pharmacokinetic and/or potency profiles. The primary 

mechanism of cardioprotection appears to be antioxidant activity: potent direct 

scavenging of the lynchpin radical in ischemia-reperfusion injury, superoxide 

anion, has been documented in appropriate model systems. In addition, 

modulation of serum complement activity, reduction of the levels of deposition of 

C-reactive protein (CRP) and the membrane attack complex (MAC) in infarcted 

tissue, and reduction in oxidative stress markers from the arachidonic acid and 

linoleic acid pathways also suggest a significant anti-inflammatory component to 

the mechanism of cardioprotection. Favorable plasma protein binding has been 

demonstrated in vitro for several astaxanthin conjugates; this binding capacity 

overcomes the supramolecular assembly of the compounds that occurs in aqueous 

solution, which in itself improves the stability and shelf-life of aqueous 

formulations. Astaxanthin readily populates cardiac tissue after metabolic 

hydrolysis of both oral and parenteral administration of the astaxanthin ester 

derivates, providing a reservoir of cardioprotective agent with a significant halflife 

due to favorable ADME in mammals. Due to the well-documented safety 

profile of astaxanthin in humans, disodium disuccinate astaxanthin may well find 

clinical utility in cardiovascular applications in humans following successful 

completion of preclinical and clinical pharmacology and toxicology studies in 

animals and humans, respectively. 




158

J Cardiovasc Pharmacol. 2006;47 Suppl 1:S7-14. 

Rofecoxib increases susceptibility of human LDL and membrane 

lipids to oxidative damage: a mechanism of cardiotoxicity. 

Mason RP, Walter MF, McNulty HP, Lockwood SF, Byun J, Day CA, Jacob 

RF. 

Cardiovascular Division, Brigham and Women's Hospital, Harvard Medical 

School, Boston, MA, USA. rpmason@elucidaresearch.com 

Clinical investigations have demonstrated a relationship between the extended use 

of rofecoxib and the increased risk for atherothrombotic events. This has led to 

the removal of rofecoxib from the market and concern over the cardiovascular 

safety of other cyclooxygenase (COX)-2 selective agents. Experimental findings 

from independent laboratories now indicate that the cardiotoxicity of rofecoxib 

may not be a class effect but because of its intrinsic chemical properties. 

Specifically, rofecoxib has been shown to increase the susceptibility of human 

low-density lipoprotein and cellular membrane lipids to oxidative modification, a 

contributing factor to plaque instability and thrombus formation. Independently of 

COX-2 inhibition, rofecoxib also promoted the nonenzymatic formation of 

isoprostanes and reactive aldehydes from biologic lipids. The basis for these 

observations is that rofecoxib alters lipid structure and readily forms a reactive 

maleic anhydride in the presence of oxygen. By contrast, other selective 

(celecoxib, valdecoxib) and nonselective (naproxen, diclofenac) inhibitors did not 

influence rates of low-density lipoprotein and membrane lipid oxidation. We have 

now further confirmed these findings by demonstrating that the prooxidant 

activity of rofecoxib can be blocked by the potent antioxidant astaxanthin in 

homochiral form (all-trans 3S, 3'S). These findings provide a mechanistic 

rationale for differences in cardiovascular risk among COX-selective inhibitors 

because of their intrinsic physicochemical properties. 



159

Biol Pharm Bull. 2006 Apr;29(4):684-8. 

Antihypertensive potential and mechanism of action of astaxanthin: 

III. Antioxidant and histopathological effects in spontaneously 

hypertensive rats. 

Hussein G, Goto H, Oda S, Sankawa U, Matsumoto K, Watanabe H. 

International Research Center for Traditional Medicine, Toyama Prefecture, 

Japan. ghazihussein@hotmail.com 

We investigated the effects of a dietary astaxanthin (ASX-O) on oxidative 

parameters in spontaneously hypertensive rats (SHR), by determination of the 

level of nitric oxide (NO) end products nitrite/nitrate (NO2-/NO3-) and lipid 

peroxidation in ASX-O-treated SHR. Oral administration of the ASX-O 

significantly reduced the plasma level of NO2-/NO3- compared to the control 

vehicle (p

J Nat Prod. 2006 Mar;69(3):443-9. 

Astaxanthin, a carotenoid with potential in human health and 

nutrition. 

Hussein G, Sankawa U, Goto H, Matsumoto K, Watanabe H. 



Astaxanthin (1), a red-orange carotenoid pigment, is a powerful biological 

antioxidant that occurs naturally in a wide variety of living organisms. The potent 

antioxidant property of 1 has been implicated in its various biological activities 

demonstrated in both experimental animals and clinical studies. Compound 1 has 

considerable potential and promising applications in human health and nutrition. 

In this review, the recent scientific literature (from 2002 to 2005) is covered on 

the most significant activities of 1, including its antioxidative and antiinflammatory 

properties, its effects on cancer, diabetes, the immune system, and 

ocular health, and other related aspects. We also discuss the green microalga 

Haematococcus pluvialis, the richest source of natural 1, and its utilization in the 

promotion of human health, including the antihypertensive and neuroprotective 

potentials of 1, emphasizing our experimental data on the effects of dietary 

astaxanthin on blood pressure, stroke, and vascular dementia in animal models, is 

described. 




161

Life Sci. 2006 Jun 6;79(2):162-74. Epub 2006 Feb 8. 

The effects of oral Cardax (disodium disuccinate astaxanthin) on multiple 

independent oxidative stress markers in a mouse peritoneal inflammation model: 

influence on 5-lipoxygenase in vitro and in vivo. 

Lockwood SF, Penn MS, Hazen SL, Bikádi Z, Zsila F. 

Hawaii Biotech, Inc., 99-193 Aiea Heights Drive, Suite 200, Aiea, Hawaii 96701, USA. 


Disodium disuccinate astaxanthin ('rac'-dAST; Cardax) is a water-dispersible C40 

carotenoid derivative under development for oral and parenteral administration for 

cardioprotection of the at-risk ischemic cardiovascular patient. In experimental infarction 

models in animals (rats, rabbits, and dogs), significant myocardial salvage has been 

obtained, up to 100% at the appropriate dose in dogs. The documented mechanism of 

action in vitro includes direct scavenging of biologically produced superoxide anion; in 

vivo in rabbits, modulation of the complement activity of serum has also been shown. A 

direct correlation between administration of the test compound in animals and reductions 

of multiple, independent markers of oxidative stress in serum was recently obtained in a 

rat experimental infarction model. For the current study, it was hypothesized that oral 

Cardax administration would inhibit oxidative damage of multiple relevant biological 

targets in a representative, well-characterized murine peritoneal inflammation model. A 

previously developed mass spectrometry-based (LC/ESI/MS/MS) approach was used to 

interrogate multiple distinct pathways of oxidation in a black mouse (C57/BL6) model 

system. In vivo markers of oxidant stress from peritoneal lavage samples (supernatants) 

were evaluated in mice on day eight (8) after treatment with either Cardax or vehicle 

(lipophilic emulsion without drug) orally by gavage at 500 mg/kg once per day for seven 

(7) days at five (5) time points: (1) baseline prior to treatment (t=0); (2) 16 h following 

intraperitoneal (i.p.) injection with thioglycollate to elicit a neutrophilic infiltrate; (3) 4 h 

following i.p. injection of yeast cell wall (zymosan; t=16 h/4 h thioglycollate+zymosan); 

(4) 72 h following i.p. injection with thioglycollate to elicit monocyte/macrophage 

infiltration; and (5) 72 h/4 h thioglycollate+zymosan. A statistically significant sparing 

effect on the arachidonic acid (AA) and linoleic acid (LA) substrates was observed at 

time points two and five. When normalized to the concentration of the oxidative 

substrates, statistically significant reductions of 8-isoprostane-F(2alpha) (8-iso-F(2alpha)) 

at time point three (maximal neutrophil recruitment/activation), and 5-HETE, 5-oxo-EET, 

11-HETE, 9-HODE, and PGF(2alpha) at time point five (maximal monocyte/macrophage 

recruitment/activation) were observed. Subsequently, the direct interaction of the 

optically inactive stereoisomer of Cardax (meso-dAST) with human 5-lipoxygenase (5- 

LOX) was evaluated in vitro with circular dichroism (CD) and electronic absorption 

(UV/Vis) spectroscopy, and subsequent molecular docking calculations were made using 

mammalian 15-LOX as a surrogate (for which XRC data has been reported). The results 

suggested that the meso-compound was capable of interaction with, and binding to, the 

solvent-exposed surface of the enzyme. These preliminary studies provide the foundation 

for more detailed evaluation of the therapeutic effects of this compound on the 5-LOX 

enzyme, important in chronic diseases such as atherosclerosis, asthma, and prostate 

cancer in humans. 



162

Mol Cell Biochem. 2006 Feb;283(1-2):23-30. 

Seven day oral supplementation with Cardax (disodium disuccinate 

astaxanthin) provides significant cardioprotection and reduces 

oxidative stress in rats. 

Gross GJ, Hazen SL, Lockwood SF. 

Department of Pharmacology and Toxicology, Medical College of Wisconsin, 

Milwaukee, 53226, USA. 

In the current study, the improved oral bioavailability of a synthetic astaxanthin 

derivative (Cardax; disodium disuccinate astaxanthin) was utilized to evaluate its 

potential effects as a cardioprotective agent after 7-day subchronic oral 

administration as a feed supplement to Sprague-Dawley rats. Animals received 

one of two concentrations of Cardax in feed (0.1 and 0.4%; approximately 125 

and 500 mg/kg/day, respectively) or control feed without drug for 7 days prior to 

the infarct study carried out on day 8. Thirty minutes of occlusion of the left 

anterior descending (LAD) coronary artery was followed by 2 h of reperfusion 

prior to sacrifice, a regimen which resulted in a mean infarct size (IS) as a 

percentage (%) of the area at risk (AAR; IS/AAR,%) of 61 +/- 1.8%. The AAR 

was quantified by Patent blue dye injection, and IS was determined by 

triphenyltetrazolium chloride (TTC) staining. Cardax at 0.1 and 0.4% in feed for 7 

days resulted in a significant mean reduction in IS/AAR,% to 45 +/- 2.0% (26% 

salvage) and 39 +/- 1.5% (36% salvage), respectively. Myocardial levels of free 

astaxanthin achieved after 7-day supplementation at each of the two 

concentrations (400 +/- 65 nM and 1634 +/- 90 nM, respectively) demonstrated 

excellent solid-tissue target organ loading after oral supplementation. Parallel 

trends in reduction of plasma levels of multiple lipid peroxidation products with 

disodium disuccinate astaxanthin supplementation were observed, consistent with 

the documented in vitro antioxidant mechanism of action. These results extend the 

potential utility of this compound for cardioprotection to the elective human 

cardiovascular patient population, for which 7-day oral pre-treatment (as with 

statins) provides significant reductions in induced periprocedural infarct size. 



163

Cardiovasc Drug Rev. 2005 Fall;23(3):199-216. 

Disodium disuccinate astaxanthin (Cardax): antioxidant and 

antiinflammatory cardioprotection. 

Lockwood SF, Gross GJ. 

Hawaii Biotech, Inc., 99-193 Aiea Heights Drive, Suite 200, Aiea, HI 96701, 

USA. slockwood@hibiotech.com 

Disodium disuccinate astaxanthin (Cardax), DDA) has cardioprotective effects in 

the rat, rabbit, and canine models of experimental infarction. It is highly effective 

by parenteral administration in subchronic and acute dosing regimens. 

Unpublished data in rats suggest that oral cardioprotection is also readily 

achievable. DDA-induced myocardial salvage in the canine can reach 100% with 

a 4-day subchronic dosing regimen. At a single i.v. dose DDA is cardioprotective, 

when given 2 h before experimental coronary occlusion, but the protection is on 

the average two-thirds of that achieved with the subchronic regimen in dogs. In 

conscious animals DDA has no effects on hemodynamic parameters. The primary 

mechanism of cardioprotection appears to be antioxidant activity involving direct 

scavenging of superoxide anion, the lynchpin radical in ischemia-reperfusion 

injury. In addition, modulation of serum complement activity, as well as the 

reduction in the levels of C-reactive protein (CRP) and the membrane attack 

complex (MAC) in infarcted tissue suggest a significant antiinflammatory 

component in the mechanism of cardioprotective action of DDA. Stoichiometric 

binding of the meso-form of the compound to human serum albumin (HSA) has 

been demonstrated in vitro. This binding capacity overcomes the supramolecular 

assembly of the compound in aqueous solution, which by itself improves the 

stability and shelf life of aqueous formulations. Non-esterified astaxanthin readily 

enters cardiac tissue after either oral or parenteral administration, providing a 

reservoir of a cardioprotective agent with a significant half-life due to favorable 

ADME in mammals. Due to the well-documented safety profile of non-esterified 

astaxanthin in humans, disodium disuccinate astaxanthin may well find clinical 

utility in cardiovascular indications in humans following successful completion of 

preclinical and clinical pharmacology and toxicology studies. 




164


Antiatherosclerotic efficacy of policosanol, red yeast rice extract and 

astaxanthin in the rabbit. 

Setnikar I, Senin P, Rovati LC. 

Rotta Research Laboratorium, Division of Rottapharm SPA, Monza, Italy. 

ivo.setnikar@rotta.com 

The effects of policosanol (P), of extract of red yeast rice (rice fermented with 

Monascus purpureus) (RYE) and of astaxanthin (A) (constituents of Armolipid) 

were investigated in a model of experimental atherosclerosis provoked in the 

rabbit by atherogenic cholesterol-enriched feed (ACEF). P and RYE and their 

combination were able to lower the increase of serum total cholesterol and of 

LDL cholesterol elicited by 3-month feeding with ACEF. They also were able to 

reduce the increase of blood malondialdehyde (MDA), a tracer of lipid 

peroxidation by the free radicals released by ACEF. When combined, the 

substances developed either additive or potentiated effects, supporting the 

rationale of their combination. Remarkable was the protective effect on lipid 

infiltration in the aortic wall provoked by ACEF, which was reduced by P and by 

RYE and almost completely prevented by the addition of A to the P-RYE 

combination. The results support the rationale of a combination of P, RYE and A 

as a useful food supplement in hyperlipemic patients. 



165

Mol Cell Biochem. 2005 Apr;272(1-2):221-7. 

Acute and chronic administration of disodium disuccinate 

astaxanthin (Cardax) produces marked cardioprotection in dog 

hearts. 

Gross GJ, Lockwood SF. 


Milwaukee, WI, USA. 

Previous results from our laboratory have shown that a novel carotenoid 

derivative (disodium disuccinate astaxanthin; Cardax) produced dose-related 

reductions in myocardial infarct size (IS) in Sprague-Dawley rats when it was 

administered at any of three doses (25, 50 and 75 mg/kg, iv) on four consecutive 

days, followed by the acute infarct size study on day 5. Maximum salvage 

occurred at the highest dose (75 mg/kg) tested, and was shown as a 56% reduction 

in IS. In the present follow-up study, we used a more relevant large animal model, 

the dog, and looked at the effect of administering Cardax iv either acutely 2 h 

prior to occlusion (N = 8) or for 4 days at 50 mg/kg iv as previously done in the 

rat model (N = 6). The results were compared to a saline vehicle-treated group (N 

= 10). In all groups, dogs were subjected to 60 min of left anterior descending 

(LAD) coronary artery occlusion and 3 h of reperfusion. IS was determined using 

a triphenyltetrazolium chloride (TTZ) histochemical stain and was expressed as a 

percent of the area at risk (IS/AAR). IS/AAR was 20.9 +/- 1.6 % (mean +/- 

S.E.M.) in controls and was reduced to 11.0 +/- 1.7% (47.3% salvage; p < 0.01) in 

dogs treated only once iv at 2 h prior to occlusion, and 6.6 +/- 2.8% (68.4% 

salvage; p < 0.001) in dogs treated for 4 days. In the chronic treatment group, two 

of the three dogs with plasma concentrations of non-esterified astaxanthin above 1 

microM had 0% IS/AAR (100% cardioprotection). These results suggest that 

Cardax has marked cardioprotective properties in both rodents and canines. Thus, 

Cardax may be a novel and powerful new means to prevent myocardial injury 

and/or necrosis associated with elective and/or urgent cardiac surgical 

interventions such as coronary angioplasty and stenting, as well as coronary artery 

bypass surgery (CABG). 



166

Biol Pharm Bull. 2005 Jun;28(6):967-71. 

Antihypertensive potential and mechanism of action of astaxanthin: 

II. Vascular reactivity and hemorheology in spontaneously 


Hussein G, Goto H, Oda S, Iguchi T, Sankawa U, Matsumoto K, Watanabe 

H. 



The current study was designed to determine the effects of a dietary astaxanthin 

(ASX-O) on vascular reactivity in spontaneously hypertensive rats (SHR), in 

order to verify its antihypertensive action mechanism. We evaluated contractions 

induced by phenylephrine (Phe), angiotensin II (Ang II) and the xanthine/xanthine 

oxidase (Xan/XOD) system, and relaxations induced by sodium nitroprusside 

(SNP) as well as endothelium-dependent relaxations mediated by acetylcholine 

(ACh) in thoracic aorta of the SHR, with and without ASX-O intervention. We 

also investigated the effects of ASX-O on blood rheology using a microchannel 

array system. In this study, ASX-O showed a significant modulatory effect on 

nitric oxide (NO)-induced vasorelaxation by the NO-donor SNP (p

J Pharmacol Exp Ther. 2005 Aug;314(2):686-92. Epub 2005 May 4. 

Disodium Disuccinate Astaxanthin (Cardax) attenuates complement 

activation and reduces myocardial injury following 

ischemia/reperfusion. 

Lauver DA, Lockwood SF, Lucchesi BR. 

Department of Pharmacology, University of Michigan Medical School, 1301C 

MSRB III, 1150 West Medical Center Drive, Ann Arbor, MI 48109, USA. 

Carotenoids are a naturally occurring group of compounds that possess 

antioxidant properties. Most natural carotenoids display poor aqueous solubility 

and tend to form aggregates in solution. Disodium disuccinate astaxanthin (DDA; 

Cardax) is a water-dispersible synthetic carotenoid that rapidly and preferentially 

associates with serum albumin, thereby preventing the formation of 

supramolecular complexes and facilitating its efficacy after parenteral 

administration. This study investigated the ability of DDA to reduce inflammation 

and myocardial injury in a rabbit model of ischemia/reperfusion. DDA (50 

mg/kg/day) or saline was administered i.v. for 4 consecutive days before the 

initiation of the protocol for induction of myocardial ischemia/reperfusion. On the 

5th day, rabbits underwent 30 min of coronary artery occlusion, followed by a 3-h 

reperfusion period. Myocardial infarct size, as a percentage of the area at risk, was 

calculated for both groups. Infarct size was 52.5 +/- 7.5% in the vehicle-treated (n 

= 9) and 25.8 +/- 4.7% in the DDA-treated (n = 9) animals (p < 0.01 versus 

vehicle; mean myocardial salvage = 51%). To evaluate the anti-inflammatory 

effects of DDA, complement activity was assessed at the end of reperfusion using 

a red blood cell lysis assay. DDA administration significantly reduced (p < 0.01) 

the activation of the complement system in the serum. The current results, 

coupled with the well established antioxidant ability of carotenoids, suggest that 

the mechanism(s) of action by which DDA reduces the tissue damage associated 

with reperfusion injury may include both antioxidant and anticomplement 

components. 




168

Biol Pharm Bull. 2005 Jan;28(1):47-52. 

Antihypertensive and neuroprotective effects of astaxanthin in 

experimental animals. 

Hussein G, Nakamura M, Zhao Q, Iguchi T, Goto H, Sankawa U, Watanabe 

H. 




living organisms. We investigated, for the first time, antihypertensive effects of 

astaxanthin (ASX-O) in spontaneously hypertensive rats (SHR). Oral 

administration of ASX-O for 14 d induced a significant reduction in the arterial 

blood pressure (BP) in SHR but not in normotensive Wistar Kyoto (WKY) strain. 

The long-term administration of ASX-O (50 mg/kg) for 5 weeks in stroke prone 

SHR (SHR-SP) induced a significant reduction in the BP. It also delayed the 

incidence of stroke in the SHR-SP. To investigate the action mechanism of ASX- 

O, the effects on PGF(2alpha)-induced contractions of rat aorta treated with NGnitro-L-arginine 

methyl ester (L-NAME) were studied in vitro. ASX-O (1 to 10 

microM) induced vasorelaxation mediated by nitric oxide (NO). The results 

suggest that the antihypertensive effect of ASX-O may be due to a NO-related 

mechanism. ASX-O also showed significant neuroprotective effects in ischemic 

mice, presumably due to its antioxidant potential. Pretreatment of the mice with 

ASX-O significantly shortened the latency of escaping onto the platform in the 

Morris water maze learning performance test. In conclusion, these results indicate 

that astaxanthin can exert beneficial effects in protection against hypertension and 

stroke and in improving memory in vascular dementia. 




169

J Mol Cell Cardiol. 2004 Nov;37(5):969-78. 

Alpha-tocopherol and astaxanthin decrease macrophage infiltration, 

apoptosis and vulnerability in atheroma of hyperlipidaemic rabbits. 

Li W, Hellsten A, Jacobsson LS, Blomqvist HM, Olsson AG, Yuan XM. 

Division of Pathology-II, Faculty of Health Sciences, Linköping University, SE- 

581 85 Linköping, Sweden. weilli@inr.liu.se 

The composition of atherosclerotic plaques, not just macroscopical lesion size, 

has been implicated in their susceptibility to rupture and the risk of thrombus 

formation. By focusing on the quality of lipids, macrophages, apoptosis, collagen, 

metalloproteinase expression and plaque integrity, we evaluated the possible antiatherosclerotic 

effect of the antioxidants alpha-tocopherol and astaxanthin in 

Watanabe heritable hyperlipidemic (WHHL) rabbits. Thirty-one WHHL rabbits 

were divided into three groups and were fed a standard diet, as controls (N =10), 

or a standard diet with the addition of 500 mg alpha-tocopherol per kg feed (N 

=11) or 100 mg astaxanthin per kg feed (N =10) for 24 weeks. We found that both 

antioxidants, particularly astaxanthin, significantly decreased macrophage 

infiltration in the plaques although they did not affect lipid accumulation. All 

lesions in the astaxanthin-treated rabbits were classified as early plaques 

according to the distribution of collagen and smooth muscle cells. Both 

antioxidants also improved plaque stability and significantly diminished 

apoptosis, which mainly occurred in macrophages, matrix metalloproteinase three 

expressions and plaque ruptures. Although neither antioxidant altered the positive 

correlations between the lesion size and lipid accumulation, the lesion size and 

apoptosis were only positively correlated in the control group. Astaxanthin and 

alpha-tocopherol may improve plaque stability by decreasing macrophage 

infiltration and apoptosis in this atherosclerotic setting. Apoptosis reduction by 

alpha-tocopherol and astaxanthin may be a new anti-atherogenic property of these 

antioxidants. 




170

Life Sci. 2004 May 28;75(2):215-24. 

Cardioprotection and myocardial salvage by a disodium disuccinate 

astaxanthin derivative (Cardax). 


Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 

Watertown Plank Road, Milwaukee, WI 53226, USA. 

Cardioprotection in humans by carotenoids has been inferred from observational and 

epidemiologic studies, however, direct studies of cardioprotection and myocardial 

salvage by carotenoids are lacking. In the current study, intravenous (I.V.) pre-treatment 

with a novel carotenoid derivative (disodium disuccinate astaxanthin; Cardax) was 

evaluated as a myocardial salvage agent in a Sprague-Dawley rat infarct model. Animals 

were dosed once per day I.V. by tail vein injection for 4 days at one of 3 doses (25, 50, 

and 75 mg/kg) prior to the infarct study carried out on day 5. The results were compared 

with control animals treated with saline vehicle. Thirty (30) minutes of occlusion of the 

left anterior descending (LAD) coronary artery was followed by 2 hours of reperfusion 

prior to sacrifice, a regimen which resulted in a mean infarct size (IS) as a percent (%) of 

the area at risk (AAR) of 59 +/- 3%. Area at risk was quantified by Patent blue dye 

injection, and infarct size (IS) was determined by triphenyltetrazolium chloride (TTC) 

staining. Cardax at 50 and 75 mg/kg for 4 days resulted in a significant mean reduction in 

IS/AAR to 35 +/- 3% (41% salvage) and 26 +/- 2% (56% salvage), respectively. Infarct 

size and myocardial salvage were significantly, and linearly, correlated with plasma 

levels of non-esterified, free astaxanthin at the end of reperfusion. These results suggest 

that parenteral Cardax may find utility in those clinical applications where pre-treatment 

of patients at risk for myocardial infarction is performed. 




171

J Atheroscler Thromb. 2000;7(4):216-22. 

Inhibition of low-density lipoprotein oxidation by astaxanthin. 

Iwamoto T, Hosoda K, Hirano R, Kurata H, Matsumoto A, Miki W, 

Kamiyama M, Itakura H, Yamamoto S, Kondo K. 

National Institute of Health and Nutrition, Tokyo, Japan. 

Marine animals produce astaxanthin which is a carotenoid and antioxidant. In this 

study we determined the in vitro and ex vivo effects of astaxanthin on LDL 

oxidation. The oxidation of LDL was measured in a 1 ml reaction system 

consisting of increasing concentrations of astaxanthin (12.5, 25.0, 50.0 

microg/ml), 400 microM V-70 (2, 2'-azobis(4-methoxy-2, 4- 

dimethylvaleronitrile)), and LDL (70 microg/ml protein). Astaxanthin dose, 

dependently significantly prolonged the oxidation lag time (31.5, 45.4, 65.0 min) 

compared with the control (19.9 min). For the ex vivo study 24 volunteers (mean 

age 28.2 [SD 7.8] years) consumed astaxanthin at doses of 1.8, 3.6,14.4 and 21.6 

mg per day for 14 days. No other changes were made in the diet. Fasting venous 

blood samples were taken at days 0, +14. LDL lag time was longer (5.0, 26.2, 

42.3 and 30.7% respectively) compared with day 0 after consuming astaxanthin at 

doses of 1.8, 3.6,14.4 and 21.6 mg for 14 days compared with day 0, but there 

was no difference in oxidation of LDL between day 0 (lag time 59.9+/-7.2 min) 

and day 14 (57.2+/-6.0 min) in the control group. Our results provide evidence 

that consumption of marine animals producing astaxanthin inhibits LDL oxidation 

and possibly therefore contributes to the prevention of atherosclerosis. 




172

Biull Eksp Biol Med. 1997 Mar;123(3):285-8. 

[Astaxanthine-induced inhibition of oxidation of apolipoprotein B- 

containing lipoproteins in human blood] 

[Article in Russian] 

Kukharchuk VV, Shumaev KB, Dmitrovskiĭ AA, Cherniad'eva IF, 

Bykhovskiĭ VIa. 



173

Prog Med F0664B 0287-3648 VOL.24;NO.6;PAGE.1437-1442(2004) 

Multivitamin and Carotenoid Supplements 

ITAKURA HIROSHIGE (Dep. Life Sci., Ibaraki Chiristian Univ., JPN) 

Abstract;Vitamins are regarded as essential nutrients for health and maintain 

stable tissue environments. Vitamins and carotenoids have multiple roles both as 

participants in many important metabolic processes throughout the body and to 

counter the oxidative stress resulting from normal metabolism and daily exposure 

to environmental agents. Epidemiological studies have consistently indicated that 

the consumption of vegetables and fruits is inversely related to the incidence of 

cardiovascular and cerebrovascular diseases and cancer. Although the majority of 

vitamins and carotenoids are derived from these foods, foods of animal origin also 

contribute supplementation of these nutrients. Marine animals supply astaxanthin 

which is a carotenoid and antioxidant. We studied the effects of astaxanthin on in 

vitro and ex vivo LDL oxidation. Astaxanthin prolonged dose-dependently the 

oxidation lag time compared with the control. For the ex vivo study 24 volunteers 

consumed astaxanthin at doses of 1.8, 3.6, 14.4, 21.6 mg per day for 14 days. 

LDL lag time was longer in the groups who intaked astaxanthin compared with 

day 0, but there was no difference in oxidation of LDL in the control group. Our 

results provide evidence that consumption of marine animals producing 

astaxanthin inhibits LDL oxidation and possibly therefore contributes to the 

prevention of atherosclerosis. 


174

Review Future Cardiology www.futuremedicine.com 

Astaxanthin, oxidative stress, inflammation and cardiovascular 

disease 

Robert G Fassett & Jeff S Coombes 

It is accepted that oxidative stress and inflammation play an integral role in the 

pathophysiology of many chronic diseases including atherosclerotic 

cardiovascular disease. The xanthophyll carotenoid dietary supplement 

astaxanthin has demonstrated potential as an antioxidant and anti-inflammatory 

therapeutic agent in models of cardiovascular disease. There have been at least 

eight clinical studies conducted in over 180 humans using astaxanthin stress, 

inflammation or the cardiovascular system. There have been no adverse 

outcomes reported. Studies have demonstrated reduced markers of oxidative 

stress and inflammation and improved blood rheology. A larger number of 

experimental studies have been performed using astaxanthin. In particular, 

studies in a variety of animals using a model of myocardial ischemia and 

reperfusion have demonstrated protective effects from prior administration of 

astaxanthin both intravenously and orally. Future clinical studies and trials will 

help determine the efficacy of antioxidants such as astaxanthin on vascular 

structure, function oxidative stress and inflammation in a variety of patients at 

risk of , or with, established cardiovascular disease. These may lead to large 

intervention trials assessing cardiovascular morbidity and mortality. 


175

Biochimica et Biophysica Acta 1768 (2007) 167–174 

Differential effects of carotenoids on lipid peroxidation due to 

membrane interactions: X-ray diffraction analysis 

Hyesun P. McNulty a,⁎, Jungsoo Byun a, Samuel F. Lockwood b, 

Robert F. Jacob a, R. Preston Mason a,c 


The biological benefits of certain carotenoids may be due to their potent 

antioxidant properties attributed to specific physico-chemical interactions with 

membranes. To test this hypothesis, we measured the effects of various 

carotenoids on rates of lipid peroxidation and correlated these findings with their 

membrane interactions, as determined by small angle X-ray diffraction 

approaches. The effects of the homochiral 

carotenoids (astaxanthin, zeaxanthin, lutein, β-carotene, lycopene) on lipid 

hydroperoxide (LOOH) generation were evaluated in membranes enriched with 

polyunsaturated fatty acids. Apolar carotenoids, such as lycopene and β-carotene, 

disordered the membrane bilayer and showed a potent pro-oxidant effect (>85% 

increase in LOOH levels) while astaxanthin preserved membrane structure and 

exhibited significant antioxidant 

activity (40% decrease in LOOH levels). These findings indicate distinct effects 

of carotenoids on lipid peroxidation due to membrane structure 

changes. These contrasting effects of carotenoids on lipid peroxidation may 

explain differences in their biological activity. 

© 2006 Elsevier B.V. All rights reserved. 


176

Antioxid Redox Signal. 2003 Feb;5(1):139-44. 

Astaxanthin limits exercise-induced skeletal and cardiac muscle 

damage in mice. 

Aoi, et al, 2003 

Dietary antioxidants may attenuate oxidative damage from strenuous exercise in 

various tissues. Beneficial effects of the antioxidant astaxanthin have been 

demonstrated in vitro, but not yet in vivo. We investigated the effect of dietary 

supplementation with astaxanthin on oxidative damage induced by strenuous 

exercise in mouse gastrocnemius and heart. C57BL/6 mice (7 weeks old) were 

divided into groups: rested control, intense exercise, and exercise with astaxanthin 

supplementation. After 3 weeks of exercise acclimation, both exercise groups ran 

on a treadmill at 28 m/min until exhaustion. Exercise-increased 4-hydroxy-2- 

nonenal-modified protein and 8-hydroxy-2'-deoxyguanosine in gastrocnemius and 

heart were blunted in the astaxanthin group. Increases in plasma creatine kinase 

activity, and in myeloperoxidase activity in gastrocnemius and heart, also were 

lessened by astaxanthin. Astaxanthin showed accumulation in gastrocnemius and 

heart from the 3 week supplementation. Astaxanthin can attenuate exerciseinduced 

damage in mouse skeletal muscle and heart, including an associated 

neutrophil infiltration that induces further damage. 


177

Mol Cell Biochem. 2005 Apr;272(1-2):221-7. 

Acute and chronic administration of disodium disuccinate 

astaxanthin (Cardax) produces marked cardioprotection in dog 

hearts. 



Milwaukee, WI, USA. 

Previous results from our laboratory have shown that a novel carotenoid 

derivative (disodium disuccinate astaxanthin; Cardax) produced dose-related 

reductions in myocardial infarct size (IS) in Sprague-Dawley rats when it was 

administered at any of three doses (25, 50 and 75 mg/kg, iv) on four consecutive 

days, followed by the acute infarct size study on day 5. Maximum salvage 

occurred at the highest dose (75 mg/kg) tested, and was shown as a 56% reduction 

in IS. In the present follow-up study, we used a more relevant large animal model, 

the dog, and looked at the effect of administering Cardax iv either acutely 2 h 

prior to occlusion (N = 8) or for 4 days at 50 mg/kg iv as previously done in the 

rat model (N = 6). The results were compared to a saline vehicle-treated group (N 

= 10). In all groups, dogs were subjected to 60 min of left anterior descending 

(LAD) coronary artery occlusion and 3 h of reperfusion. IS was determined using 

a triphenyltetrazolium chloride (TTZ) histochemical stain and was expressed as a 

percent of the area at risk (IS/AAR). IS/AAR was 20.9 +/- 1.6 % (mean +/- 

S.E.M.) in controls and was reduced to 11.0 +/- 1.7% (47.3% salvage; p < 0.01) in 

dogs treated only once iv at 2 h prior to occlusion, and 6.6 +/- 2.8% (68.4% 

salvage; p < 0.001) in dogs treated for 4 days. In the chronic treatment group, two 

of the three dogs with plasma concentrations of non-esterified astaxanthin above 1 

microM had 0% IS/AAR (100% cardioprotection). These results suggest that 

Cardax has marked cardioprotective properties in both rodents and canines. Thus, 

Cardax may be a novel and powerful new means to prevent myocardial injury 

and/or necrosis associated with elective and/or urgent cardiac surgical 

interventions such as coronary angioplasty and stenting, as well as coronary artery 

bypass surgery (CABG). 



178

Biosci Biotechnol Biochem. 2007 Apr;71(4):893-9. Epub 2007 Apr 7. 

Effects of astaxanthin in obese mice fed a high-fat diet. 

Ikeuchi M, Koyama T, Takahashi J, Yazawa K. 

Laboratory of Nertraceuticals and Functional Foods Science, Graduate School of 

Marine Science and Technology, Tokyo University of Marine Science and 

Technology, Tokyo, Japan. 


living organisms. We investigated the effects of astaxanthin supplementation in 

obese mice fed a high-fat diet. Astaxanthin inhibited the increases in body weight 

and weight of adipose tissue that result from feeding a high-fat diet. In addition, 

astaxanthin reduced liver weight, liver triglyceride, plasma triglyceride, and total 

cholesterol. These results suggest that astaxanthin might be of value in reducing 

the likelihood of obesity and metabolic syndrome in affluent societies. 


179

Organo Oficial de la Sociedad Latinoamericana de Nutricion Vol.42 No4. 1992 

The Effect of Canthaxanthin and Astaxanthin on 

Hypercholesterolemic on Rats 

Enrique Murillo 

Hypercholesterolemic effects of canthaxanthin and astaxanthin in rats. Three 

groups of male Wistar rats (130-140 g) were fed 30 days with a synthetic diets 

containing 0,1% of β-carotene, canthaxanthin and astaxanthin respectively. 

Another group was fed with a synthetic diet without carotenoids. The results 

shows that the β-carotene dies not induce change in plasma cholesterol (49, 7±3,6 

mg/dl), but canthaxanthin and astaxanthin induce a significant increase in 

cholesterol concentration (92,1±3,6 and 66,1±5,1 mg/dl). This increase is noted 

mainly in the HDL fraction of the lipoproteins. Canthaxanthin has more affinity 

than astaxanthin for the liver, principal site of lipoproteins catabolism. The 

hipercholesterolemic effect of these xanthophylls is not related to reported 

mechanisms of carotenoids in mammalian, because β-carotene does not induce 

changes in plasma cholesterol. 


180

Biol Pharm Bull S0989A 0918-6158 VOL.29;NO.4;PAGE.684-688 (J-STAGE)(2006) 

Antihypertensive Potential and Mechanism of Action of 

Astaxanthin: III. Antioxidant and Histopathological Effects in 

Spontaneously Hypertensive Rats 

HUSSEIN GHAZI HUSSEIN GHAZI GOTO HIROZO ODA SHINOBU 

SANKAWA USHIO MATSUMOTO KINZO (WATANABE HIROSHI 

Abstract;We investigated the effects of a dietary astaxanthin (ASX-O) on 

oxidative parameters in spontaneously hypertensive rats (SHR), by determination 

of the level of nitric oxide (NO) end products nitrite/nitrate (NO2'-'/NO3'-') and 

lipid peroxidation in ASX-O-treated SHR. Oral administration of the ASX-O 

significantly reduced the plasma level of NO2'-'/NO3'-' compared to the control 

vehicle (p

Biol Pharm Bull S0989A 0918-6158 VOL.28;NO.6;PAGE.967-971(2005) 

Antihypertensive Potential and Mechanism of Action of 

Astaxanthin: II. Vascular Reactivity and Hemorheology in 

Spontaneously Hypertensive Rats 

HUSSEIN GHAZI GOTO HIROZO ODA SHINOBU IGUCHI TOMOMI 

SANKAWA USHIO MATSUMOTO KINZO WATANABE HIROSHI 

Abstract;The current study was designed to determine the effects of a dietary 

astaxanthin (ASX-O) on vascular reactivity in spontaneously hypertensive rats 

(SHR), in order to verify its antihypertensive action mechanism. We evaluated 

contractions induced by phenylephrine (Phe), angiotensin II (Ang II) and the 

xanthine/xanthine oxidase (Xan/XOD) system, and relaxations induced by sodium 

nitroprusside (SNP) as well as endothelium-dependent relaxations mediated by 

acetylcholine (ACh) in thoracic aorta of the SHR, with and without ASX-O 

intervention. We also investigated the effects of ASX-O on blood rheology using 

a microchannel array system. In this study, ASX-O showed a significant 

modulatory effect on nitric oxide (NO)-induced vasorelaxation by the NO-donor 

SNP (p

JOURNAL OF FUNCTIONAL FOODS 1 (2009) 13–22 

Astaxanthin lowers blood pressure and lessens the activity of the 

renin-angiotensin system in Zucker Fatty Rats 

Harry G. Preussa,*, Bobby Echarda, Debasis Bagchib, Nicholas V. Perriconec, 

Eiji Yamashita 

The ability of astaxanthin to favorably influence the renin-angiotensin system 

(RAS), blood pressure (BP), and metabolic parameters in Zucker Fatty Rats 

(ZFR) was examined. In separate experiments, 96 ZFR were equally divided into 

four groups: control, captopril (30 mg/kg), low astaxanthin (5 mg/kg) and high 

astaxanthin (25 mg/kg). RAS and insulin systems were examined following 

recovery from heat stress. RAS was lower in test groups; however, there was no 

evidence of enhanced insulin sensitivity. Test groups decreased SBP (systolic 

blood pressure) significantly compared to the control. The tests carried out 

suggested that RAS was involved in the ability of astaxanthin to lower BP. 

Astaxanthin at high dosage influenced circulating TNF-a and MCP-1 and lessened 

fat oxidation in liver and kidneys. Thus, astaxanthin may be considered as a good 

stress reducer with regards to heat stress. Astaxanthin’s effects on RAS indicate it 

might overcome perturbations associated with increased activity, especially those 

related to the cardiovascular system. 


183

J Mol Cell Cardiol. 2004 Nov;37(5):969-78. 

Alpha-tocopherol and astaxanthin decrease macrophage infiltration, 

apoptosis and vulnerability in atheroma of hyperlipidaemic rabbits. 

The composition of atherosclerotic plaques, not just macroscopical lesion size, 

has been implicated in their susceptibility to rupture and the risk of thrombus 

formation. By focusing on the quality of lipids, macrophages, apoptosis, collagen, 

metalloproteinase expression and plaque integrity, we evaluated the possible antiatherosclerotic 

effect of the antioxidants alpha-tocopherol and astaxanthin in 

Watanabe heritable hyperlipidemic (WHHL) rabbits. Thirty-one WHHL rabbits 

were divided into three groups and were fed a standard diet, as controls (N =10), 

or a standard diet with the addition of 500 mg alpha-tocopherol per kg feed (N 

=11) or 100 mg astaxanthin per kg feed (N =10) for 24 weeks. We found that both 

antioxidants, particularly astaxanthin, significantly decreased macrophage 

infiltration in the plaques although they did not affect lipid accumulation. All 

lesions in the astaxanthin-treated rabbits were classified as early plaques 

according to the distribution of collagen and smooth muscle cells. Both 

antioxidants also improved plaque stability and significantly diminished 

apoptosis, which mainly occurred in macrophages, matrix metalloproteinase three 

expressions and plaque ruptures. Although neither antioxidant altered the positive 

correlations between the lesion size and lipid accumulation, the lesion size and 

apoptosis were only positively correlated in the control group. Astaxanthin and 

alpha-tocopherol may improve plaque stability by decreasing macrophage 

infiltration and apoptosis in this atherosclerotic setting. Apoptosis reduction by 

alpha-tocopherol and astaxanthin may be a new anti-atherogenic property of these 

antioxidants. 


184

J of the Pharmaceutical Society of Japan VOL.126;NO.Suppl.3;PAGE.16-19(2006) 

PREVENTION BY ASTAXANTHIN OF LIFE STYLE DISEASES: 

EXPERIMENTAL EVIDENCES 

WATANABE HIROSHI; HUSSEIN GHAZI; GOTO HIROZO; NAKAGAWA 

TAKAKO; MATSUMOTO KINZO; SANKAWA USHIO 

Astaxanthin (ASX), a red-orange carotenoid pigment, is a powerful antioxidant 

that occurs naturally in a wide variety of living organisms. We investigated the 

effect of ASX on the incidence of stroke, hypertension, and hyperglycemia in rats. 

Repeated ASX (50 mg/kg/day, p.o.) inhibited the incidence of stroke in SHRstroke 

prone (SP). Pretreatment with 50 mg/kg/day of ASX for a week produced 

anti-hypertensive effect in awaked SHR. In the isolated aorta, ASX inhibited the 

vascular contraction induced by PGF2.ALPHA.. Pretreatment with L-NAME 

(10'-4'M) ameliorated the inhibitory effect of ASX. ASX produced a significant 

reduction in the elastin bands and diminished the wall thickness in the SHR aorta. 

Fifty mg/kg of ASX for 18 weeks caused a significant decrease in the blood 

glucose in SHR/ND mcr-cp (cp/cp). ASX (50 mg/kg) produced a tendency to 

improve the learning behavior deficit induced by the brain ischemia in mice. 

These results suggest that ASX may exert beneficial effects for the protection 

against lifestyle related diseases. 


185

J Cardiovasc Pharmacol. 2006 May;47 Suppl 1:S7-S14. 

Related Articles, Links 

Rofecoxib Increases Susceptibility of Human LDL and Membrane 

Lipids to Oxidative Damage: A Mechanism of Cardiotoxicity. 

Preston Mason R, Walter MF, McNulty HP, Lockwood SF, Byun J, Day CA, 

Jacob RF. 

*Cardiovascular Division, Brigham and Women's Hospital, Harvard Medical 

School, Boston, MA daggerElucida Research LLC, Beverly, MA double 

daggerAtlanta VA Medical Center, Atlanta, GA section signCardax 

Pharmaceuticals, Inc, Aiea, HI. 

Clinical investigations have demonstrated a relationship between the extended use 

of rofecoxib and the increased risk for atherothrombotic events. This has led to 

the removal of rofecoxib from the market and concern over the cardiovascular 

safety of other cyclooxygenase (COX)-2 selective agents. Experimental findings 

from independent laboratories now indicate that the cardiotoxicity of rofecoxib 

may not be a class effect but because of its intrinsic chemical properties. 

Specifically, rofecoxib has been shown to increase the susceptibility of human 

low-density lipoprotein and cellular membrane lipids to oxidative modification, a 

contributing factor to plaque instability and thrombus formation. Independently of 

COX-2 inhibition, rofecoxib also promoted the nonenzymatic formation of 

isoprostanes and reactive aldehydes from biologic lipids. The basis for these 

observations is that rofecoxib alters lipid structure and readily forms a reactive 

maleic anhydride in the presence of oxygen. By contrast, other selective 

(celecoxib, valdecoxib) and nonselective (naproxen, diclofenac) inhibitors did not 

influence rates of low-density lipoprotein and membrane lipid oxidation. We have 

now further confirmed these findings by demonstrating that the prooxidant 

activity of rofecoxib can be blocked by the potent antioxidant astaxanthin in 

homochiral form (all-trans 3S, 3'S). These findings provide a mechanistic 

rationale for differences in cardiovascular risk among COX-selective inhibitors 

because of their intrinsic physicochemical properties. 


186

Pharmacol Res. 2010 Sep 22. [Epub ahead of print] 

Astaxanthin-enriched-diet reduces blood pressure and improves 

cardiovascular parameters in spontaneously hypertensive rats. 

Monroy-Ruiz J, Sevilla MA, Carrón R, Montero MJ. 

Departamento de Salud, Universidad Iberoamericana, Prolongación Paseo de la Reforma 

880, Ciudad de México, Mexico. 


The aim of this study was to investigate the effects of astaxanthin-enriched diet on blood 

pressure, cardiac hypertrophy, both vascular structure and function and superoxide 

(O(2)(-)) production in spontaneously hypertensive rats (SHR). Twelve-week-old SHR 

were treated for 8 weeks with an astaxanthin-enriched diet (75 or 200mg/kg body weight 

per day). Systolic blood pressure was monitorized periodically during the study by the 

tail cuff method. At the end of the study animals were sacrificed and heart, kidneys and 

aorta were removed. Left ventricular weight/body weight ratio was used as left 

ventricular hypertrophy index (LVH). Vascular function and structure were studied in 

conductance (aortic rings) and resistance (renal vascular bed) arteries. Also O(2)(-) 

production was evaluated by lucigenin-enhanced chemiluminescence. Systolic blood 

pressure was lower in astaxanthin-treated groups than the control group from the first 

week of treatment, and LVH was significantly reduced. Astaxanthin improved 

endothelial function on resistance arteries, but had no effect on aorta. These effects were 

accompanied by a decrease in oxidative stress and improvements in NO bioavailability. 

Taken together, these results show that diet supplemented with astaxanthin has beneficial 

effects on hypertension, by decreasing blood pressure values, improving cardiovascular 

remodeling and oxidative stress. 



187

Future Cardiol. 2009 Jul;5(4):333-42. 

Astaxanthin, oxidative stress, inflammation and cardiovascular disease. 

Fassett RG, Coombes JS. 

School of Human Movement Studies & School of Medicine, The University of 

Queensland, Queensland, Australia. r.fassett@uq.edu.au 


It is accepted that oxidative stress and inflammation play an integral role in the 

pathophysiology of many chronic diseases including atherosclerotic cardiovascular 

disease. The xanthophyll carotenoid dietary supplement astaxanthin has demonstrated 

potential as an antioxidant and anti-inflammatory therapeutic agent in models of 

cardiovascular disease. There have been at least eight clinical studies conducted in over 

180 humans using astaxanthin to assess its safety, bioavailability and clinical aspects 

relevant to oxidative stress, inflammation or the cardiovascular system. There have been 

no adverse outcomes reported. Studies have demonstrated reduced markers of oxidative 

stress and inflammation and improved blood rheology. A larger number of experimental 

studies have been performed using astaxanthin. In particular, studies in a variety of 

animals using a model of myocardial ischemia and reperfusion have demonstrated 

protective effects from prior administration of astaxanthin both intravenously and orally. 

Future clinical studies and trials will help determine the efficacy of antioxidants such as 

astaxanthin on vascular structure, function, oxidative stress and inflammation in a variety 

of patients at risk of, or with, established cardiovascular disease. These may lead to large 

intervention trials assessing cardiovascular morbidity and mortality. 



188

Eur J Nutr. 2010 Mar;49(2):119-26. Epub 2009 Sep 26. 

Astaxanthin suppresses scavenger receptor expression and matrix 

metalloproteinase activity in macrophages. 

Kishimoto Y, Tani M, Uto-Kondo H, Iizuka M, Saita E, Sone H, Kurata H, Kondo K. 

Institute of Environmental Science for Human Life, Ochanomizu University, Tokyo, 

Japan. 


BACKGROUND: Astaxanthin is a red carotenoid pigment which has significant 

potential for antioxidant activity. The macrophages in atherosclerotic lesions, known as 

activated macrophages, express scavenger receptors responsible for the clearance of 

pathogenic lipoproteins. In addition, the expression and secretion of proteolytic enzymes, 

matrix metalloproteinases (MMPs), and pro-inflammatory cytokines are remarkably 

promoted in activated macrophages. 

AIM OF THE STUDY: In this study, we investigated the effects of astaxanthin on the 

expression of scavenger receptors, MMPs, and pro-inflammatory cytokines in 

macrophages. 

METHODS: THP-1 macrophages were incubated with 5-10 microM astaxanthin for 24 

h. The expression levels of scavenger receptors, MMPs, and pro-inflammatory cytokines 

were determined by Western blot analysis or real-time RT-PCR. The MMP-9 and -2 

activities were examined by gelatin zymography and total MMP activity was measured 

by fluorometry. 

RESULTS: We found that astaxanthin remarkably decreased the class A scavenger 

receptor and CD36 expression in the protein and mRNA levels. Astaxanthin also reduced 

MMP-1, -2, -3, -9, -12, and -14 activity and expression. The mRNA expression of tumor 

necrosis factor-alpha, interleukin-1beta, interleukin-6, inducible nitric oxide synthase, 

and cyclooxygenase-2 were significantly suppressed by astaxanthin. Furthermore, 

astaxanthin inhibited the phosphorylation of nuclear factor-kappaB. 

189

CONCLUSIONS: These results indicate that astaxanthin has inhibitory effects on 

macrophage activation, such as scavenger receptors up-regulation, MMPs activation, and 

pro-inflammatory cytokines secretion. 



190

Thromb Res. 2010 Oct;126(4):299-305. Epub 2010 Aug 21. 

Novel astaxanthin prodrug (CDX-085) attenuates thrombosis in a 

mouse model. 

Khan SK, Malinski T, Mason RP, Kubant R, Jacob RF, Fujioka K, Denstaedt SJ, King 

TJ, Jackson HL, Hieber AD, Lockwood SF, Goodin TH, Pashkow FJ, Bodary PF. 

School of Kinesiology, University of Michigan, Ann Arbor, MI, USA. 


BACKGROUND: Cardiovascular disease remains the leading cause of morbidity and 

premature mortality in most industrialized countries as well as in developing nations. A 

pro-oxidative state appears to promote and/or exacerbate vascular disease complications. 

Furthermore, a state of low-grade chronic inflammation can promote increased oxidative 

stress and lead to endothelial cell and platelet dysfunction ultimately contributing to 

thrombogenesis. 

OBJECTIVES: In this study, the effect of a proprietary astaxanthin prodrug (CDX-085) 

on thrombus formation was investigated using a mouse model of arterial thrombosis. The 

influence of free astaxanthin, the active drug of CDX-085, on human endothelial cells 

and rat platelets was evaluated to investigate potential mechanisms of action. 

METHODS AND RESULTS: Oral administration of CDX-085 (0.4% in chow, 

approximately 500 mg/kg/day) to 6-8 week old C57BL/6 male mice for 14 days resulted 

in significant levels of free astaxanthin in the plasma, liver, heart and platelets. When 

compared to control mice, the CDX-085 fed group exhibited significant increases in basal 

arterial blood flow and significant delays in occlusive thrombus formation following the 

onset of vascular endothelial injury. Primary human umbilical vein endothelial cells 

(HUVECs) and platelets isolated from Wistar-Kyoto rats treated with free astaxanthin 

demonstrated significantly increased levels of released nitric oxide (NO) and significantly 

decreased peroxynitrite (ONOO-) levels. 

CONCLUSION: Observations of increased NO and decreased ONOO- levels in 

endothelial cells and platelets support a potential mechanism of action for astaxanthin 

(CDX-085 active drug). These studies support the potential of CDX-085 and its 

metabolite astaxanthin in the treatment or prevention of thrombotic cardiovascular 

complications. 



191

Anticancer Res. 2010 Jul;30(7):2721-5. 

Effect of astaxanthin supplementation on inflammation and cardiac 

function in BALB/c mice. 

Nakao R, Nelson OL, Park JS, Mathison BD, Thompson PA, Chew BP. 

School of Food Science, Washington State University, Pullman, WA 99164, USA. 


Astaxanthin is an antioxidant with immunomodulatory, anti-inflammatory and anticancer 

properties. This study evaluated the use of dietary astaxanthin to decrease oxidative stress 

and improve cardiac function, thereby providing a potential cardioprotective supplement. 

Female BALB/c mice (8 weeks of age) were fed a semi-synthetic diet containing 0, 0.02 

or 0.08% astaxanthin for 8 weeks. Cardiac function was assessed by echocardiography 

bi-weekly, and blood and tissue samples were collected at 8 weeks. Plasma astaxanthin 

concentrations increased (p

J Cardiovasc Pharmacol Ther. 2009 Dec;14(4):314-22. Epub 2009 Oct 21. 

Astaxanthin reduces oxidative stress, but not aortic damage in 

atherosclerotic rabbits. 

Augusti PR, Conterato GM, Somacal S, Sobieski R, Quatrin A, Maurer L, Rocha MP, 

Denardin IT, Emanuelli T. 

Department of Biochemistry, Institute of Health Basic Sciences, Federal University of 

Rio Grande do Sul, Porto Alegre, RS, Brazil. 


We evaluated whether carotenoid astaxanthin (ASX) could prevent oxidative and 

atherosclerotic damage in rabbits. Rabbits received regular chow (control) or an 

atherogenic diet (1% cholesterol) alone or supplemented with 50, 100, and 500 mg% 

ASX for 60 days (n = 5-9 per group). The atherogenic diet increased the serum 

cholesterol levels and the ratio of the intima/media area in the aortic arch. These changes 

were not prevented by ASX. Atherosclerotic rabbits showed increased aortic lipid 

peroxidation and nonprotein thiol group (NPSH) levels along with inhibition of 

glutathione peroxidase (GSH-Px). All ASX doses attenuated lipid peroxidation and the 

increase in NPSH but not the inhibition of GSH-Px. Aortic superoxide dismutase (SOD), 

catalase (CAT), and thioredoxin reductase (TrxR) activities were enhanced in 

atherosclerotic rabbits. Although all ASX doses prevented the increase in SOD activity, 

only 100 and 500 mg% ASX prevented the increase in CAT activity. Furthermore, these 

same doses partially prevented the increase in TrxR activity, while 50 mg% ASX 

completely prevented the effects of the atherogenic diet on this enzyme. However, ASX 

did not attenuate the hypercholesterolemia or the atherosclerotic lesions caused by the 

atherogenic diet at any of the doses evaluated. Our results indicate that although ASX did 

not prevent hypercholesterolemia or atherosclerotic lesions, it could play a beneficial role 

by preventing lipid peroxidation and changes in antioxidant enzyme activities. 



193

Curr Atheroscler Rep. 2009 Nov;11(6):434-9. 

Carotenoids and cardiovascular disease. 

Riccioni G. 

Cardiology Unit, San Camillo de Lellis Hospital, Manfredonia (FG), Italy. 

griccioni@hotmail.com 


Carotenoids are a class of natural fat-soluble pigments found principally in plants. They 

have potential antioxidant biological properties due to their chemical structure and 

interaction with biological membranes. The most abundant carotenoids in the diet are 

beta-carotene, lycopene, lutein, beta-cryptoxanthin, zeaxanthin, and astaxanthin. 

Numerous epidemiologic studies have supported the hypothesis that antioxidants could 

be used as an inexpensive means of prevention, and possibly treatment, of cardiovascular 

diseases, even though findings from interventional trials have been mixed, with some 

positive findings, many null findings, and some suggestion of harm in certain high-risk 

populations. Recent smaller interventional studies with carefully chosen populations, 

such as those under high levels of oxidative stress, have yielded largely positive results. 

This suggests that we need more hypothesis-driven and rigorous clinical trial designs. 

The aim of this review is to examine the published studies about the use of carotenoids, 

especially lycopene and astaxanthin, in the treatment of cardiovascular diseases. 



194

Atherosclerosis. 2010 Apr;209(2):520-3. Epub 2009 Oct 14. 

Administration of natural astaxanthin increases serum 

HDL-cholesterol and adiponectin in subjects with mild 

hyperlipidemia. 

Yoshida H, Yanai H, Ito K, Tomono Y, Koikeda T, Tsukahara H, Tada N. 

Source 

Department of Laboratory Medicine, Jikei University Kashiwa Hospital, Chiba, Japan. 

hyoshida@jikei.ac.jp 


BACKGROUND: Astaxanthin has been reported to improve dyslipidemia and metabolic 

syndrome in animals, but such effects in humans are not well known. 

METHODS: Placebo-controlled astaxanthin administration at doses of 0, 6, 12, 18 

mg/day for 12 weeks was randomly allocated to 61 non-obese subjects with fasting serum 

triglyceride of 120-200mg/dl and without diabetes and hypertension, aged 25-60 years. 

RESULTS: In before and after tests, body mass index (BMI) and LDL-cholesterol were 

unaffected at all doses, however, triglyceride decreased, while HDL-cholesterol increased 

significantly. Multiple comparison tests showed that 12 and 18 mg/day doses 

significantly reduced triglyceride, and 6 and 12 mg doses significantly increased HDLcholesterol. 

Serum adiponectin was increased by astaxanthin (12 and 18 mg/day), and 

changes of adiponectin correlated positively with HDL-cholesterol changes independent 

of age and BMI. 

CONCLUSIONS: This first-ever randomized, placebo-controlled human study suggests 

that astaxanthin consumption ameliorates triglyceride and HDL-cholesterol in correlation 

with increased adiponectin in humans. 

Copyright 2009 Elsevier Ireland Ltd. All rights reserved. 



195

Pharmacol Res. 2011 Jan;63(1):44-50. Epub 2010 Sep 22. 

Astaxanthin-enriched-diet reduces blood pressure and 

improves cardiovascular parameters in spontaneously 


Monroy-Ruiz J, Sevilla MÁ, Carrón R, Montero MJ. 

Source 

Departamento de Salud, Universidad Iberoamericana, Prolongación Paseo de la Reforma 

880, Ciudad de México, Mexico. 


The aim of this study was to investigate the effects of astaxanthin-enriched diet on blood 

pressure, cardiac hypertrophy, both vascular structure and function and superoxide 

((*)O(2-)) production in spontaneously hypertensive rats (SHR). Twelve-week-old SHR 

were treated for 8 weeks with an astaxanthin-enriched diet (75 or 200mg/kg body weight 

per day). Systolic blood pressure was monitorized periodically during the study by the 

tail cuff method. At the end of the study animals were sacrificed and heart, kidneys and 

aorta were removed. Left ventricular weight/body weight ratio was used as left 

ventricular hypertrophy index (LVH). Vascular function and structure were studied in 

conductance (aortic rings) and resistance (renal vascular bed) arteries. Also (*)O(2-) 

production was evaluated by lucigenin-enhanced chemiluminescence. Systolic blood 

pressure was lower in astaxanthin-treated groups than the control group from the first 

week of treatment, and LVH was significantly reduced. Astaxanthin improved 

endothelial function on resistance arteries, but had no effect on aorta. These effects were 

accompanied by a decrease in oxidative stress and improvements in NO bioavailability. 

Taken together, these results show that diet supplemented with astaxanthin has beneficial 

effects on hypertension, by decreasing blood pressure values, improving cardiovascular 

remodeling and oxidative stress. 

Copyright Â© 2010 Elsevier Ltd. All rights reserved. 



196

Integr Blood Press Control. 2008;1:1-3. Epub 2008 Oct 27. 

Antihypertensive effects of astaxanthin. 

Yanai H, Ito K, Yoshida H, Tada N. 

Source 

Department of Internal Medicine; 


Astaxanthin is a biological antioxidant naturally found in a wide variety of aquatic living 

organisms, and has shown various pharmacological activities, such as anti-inflammatory 

and antidiabetic activities. A recent study reported that the administration of astaxanthin 

induced a significant reduction in blood pressure and delayed the incidence of stroke in 

stroke-prone spontaneously hypertensive rats, suggesting that astaxanthin also has 

antihypertensive effect. In a study using aortic rings of spontaneously hypertensive rats, 

astaxanthin induced a significant reduction of the contractile responses of the aorta to α- 

adrenergic receptor agonist and angiotensin II, which may contribute to the 

antihypertensive effect of astaxanthin. In a histopathological study, astaxanthin decreased 

coronary artery wall thickness compared with the control, indicating the possibility that 

astaxanthin ameliorates hypertension-induced vascular remodeling. Astaxanthin has antiinflammatory, 

antidiabetic, antihypertensive, and antioxidative activities; therefore, we 

should perform further studies to elucidate an antiatherogenic effect of astaxanthin. 




197

Immunity 

Nutr Metab (Lond). 2010 Mar 5;7:18. 

Astaxanthin decreased oxidative stress and inflammation and enhanced 

immune response in humans. 

Park JS, Chyun JH, Kim YK, Line LL, Chew BP. 

School of Food Science, Washington State University, Pullman, WA 99164-6376 USA. 

boonchew@wsu.edu. 

ABSTRACT: 

BACKGROUND: Astaxanthin modulates immune response, inhibits cancer cell growth, 

reduces bacterial load and gastric inflammation, and protects against UVA-induced 

oxidative stress in in vitro and rodent models. Similar clinical studies in humans are 

unavailable. Our objective is to study the action of dietary astaxanthin in modulating 

immune response, oxidative status and inflammation in young healthy adult female 

human subjects. 

METHODS: Participants (averaged 21.5 yr) received 0, 2, or 8 mg astaxanthin (n = 

14/diet) daily for 8 wk in a randomized double-blind, placebo-controlled study. Immune 

response was assessed on wk 0, 4 and 8, and tuberculin test performed on wk 8. 

RESULTS: Plasma astaxanthin increased (P < 0.01) dose-dependently after 4 or 8 wk of 

supplementation. Astaxanthin decreased a DNA damage biomarker after 4 wk but did not 

affect lipid peroxidation. Plasma C-reactive protein concentration was lower (P < 0.05) 

on wk 8 in subjects given 2 mg astaxanthin. Dietary astaxanthin stimulated mitogeninduced 

lymphoproliferation, increased natural killer cell cytotoxic activity, and 

increased total T and B cell subpopulations, but did not influence populations of Thelper, 

Tcytotoxic or natural killer cells. A higher percentage of leukocytes expressed the LFA-1 

marker in subjects given 2 mg astaxanthin on wk 8. Subjects fed 2 mg astaxanthin had a 

higher tuberculin response than unsupplemented subjects. There was no difference in 

TNF and IL-2 concentrations, but plasma IFN-gamma and IL-6 increased on wk 8 in 

subjects given 8 mg astaxanthin. 

CONCLUSION: Therefore, dietary astaxanthin decreases a DNA damage biomarker and 

acute phase protein, and enhances immune response in young healthy females. 


Immunity 

198

Experimental Biology Saturday, April 17, 2004 

Abstract #341.4 

Immune stimulating action of dietary astaxanthin in humans 

J. Park, J. Chyun, Y. Kim, L. Line, M. Maloney and B. Chew 

We studied the role of dietary astaxanthin on immunity and oxidative status. 

Female subjects (21.5 yr) with no history of major diseases received 0, 2, or 8 mg 

astaxanthin (n = 14) daily for 8 wk in a double-blind, placebo controlled study. 

Blood was drawn on wk 0, 4 and 8. The tuberculin test was assessed on wk 8. 

Plasma astaxanthin was undetectable prior to feeding but increased (P < 0.01) 

dose-dependently on wk 4 and 8. Dietary astaxanthin stimulated concanavalin A-, 

phytohemagglutinin- and pokeweed mitogen-induced lymphoproliferation and 

increased NK cell cytotoxic activity. In addition, astaxanthin also increased the 

proportion of total T cells and B cells, but did not influence the populations of Th, 

Tc or NK cells or the ratio of Th:Tc cells. The frequency of cells expressing LFA- 

1 marker was higher in subjects given 2 mg (42.1%) but not those given 8 mg 

(30.6%) astaxanthin compare to control (31.8%) on wk 8. No similar dietary 

effect was observed with ICAM-1 or LFA-3 expression. Subjects fed 2 mg but not 

those fed 8 mg astaxanthin had higher DTH response than unsupplemented 

controls. Dietary astaxanthin dramatically decreased blood DNA damage (8- 

oxodeoxyguanosine) after 4 wk of feeding but did not influence lipid peroxidation 

in plasma. Therefore, dietary astaxanthin enhanced immune response and 

decrease DNA damage in human subjects. 

Immunity 

199

Int J Immunopharmacol. 1996 Dec;18(12):753-8. 

Possible immunomodulating activities of carotenoids in in vitro cell 

culture experiments. 

Okai Y, Higashi-Okai K. 

Division of Food and Nutrition, Osaka Kun-Ei Women's College, Japan. 

Immunomodulating activities of beta-carotene and carotene-associated 

carotenoids such as canthaxanthin (beta, beta-carotene-4,4 dione) and astaxanthin 

(3,3'-dihydroxyl beta, beta-carotene 4,4-dione) were analyzed by in vitro cell 

culture experiments. (i) beta-Carotene, canthaxanthin and astaxanthin caused 

significant stimulatory effects on the cell proliferative response of spleen cells and 

thymocytes from BALB/c mice at the concentrations of 2 x 10(-8) to 10(-7) M, 

although they showed the activities different from each other. (ii) Astaxanthin 

exhibited the highest activity on the polyclonal antibody (immunoglobulin M and 

G) production of murine spleen cells at the concentrations of 2 x 10(-8) to 10(-7) 

M but beta-carotene did not cause a significant effect at a low concentration (2 x 

10(-8) M) although stimulated at a high concentration (2 x 10(-7) M). 

Canthaxanthin expressed moderate activities at the same concentrations. (iii) All 

tested carotenoids significantly enhanced the release of interleukin-1 alpha and 

tumor necrosis factor-alpha from murine peritoneal adherent cells at the 

concentrations of 2 x 10(-8) to 10(-7) M and the ranks of cytokine-inducing 

activities were astaxanthin > canthaxanthin > beta-carotene. These results indicate 

that carotenoids such as beta-carotene, canthaxanthin and astaxanthin have 

possible immunomodulating activities to enhance the proliferation and functions 

of murine immunocompetent cells. 


Immunity 

200

Nutr Cancer. 1996;26(3):313-24. 

Effects of various carotenoids on cloned, effector-stage T-helper cell 

activity. 

Jyonouchi H, Sun S, Mizokami M, Gross MD. 

Department of Pediatrics, School of Medicine, University of Minnesota, 

Minneapolis 55455, USA. jyono001@maroon.tc.umn.edu 

Astaxanthin, a carotenoid without provitamin A activity, enhances murine T- 

helper (Th) cell clone-mediated antibody (Ab) production with suboptimal 

antigen (Ag) challenges. It also suppresses interferon-gamma (IFN-gamma) 

production by cloned murine Th1 cells. beta-Carotene is less effective than 

astaxanthin. This study evaluates the effects of various carotenoids with various 

relative polarity, provitamin A activity, and antioxidant activity. Carotenoids 

tested include astaxanthin, cantaxanthin, zeaxanthin, lutein, and lycopene, and 

their effects were tested at a concentration at which astaxanthin's effect was most 

potent. A.E7 and CDC35 cells are used as representative type 1 and type 2 Th cell 

(Th1 and Th2) clones, respectively. In the Th1 clone, astaxanthin, but not other 

carotenoids, suppressed IFN-gamma production and increased the number of Absecreting 

cells with the use of primed spleen cells. With cultures of Th1 cells and 

unprimed spleen cells, astaxanthin and zeaxanthin augmented the number of 

immunoglobulin M Ab-secreting cells. In the cultures of Th2 clone and primed 

spleen cells, astaxanthin, but not other carotenoids, enhanced the number of Absecreting 

cells. With unprimed spleen cells, lycopene suppressed Th2 clonemediated 

Ab production. Interleukin-5 production by the Th2 clone was not 

significantly altered with the carotenoids tested, irrespective of the use of 

unprimed or primed spleen cells. Carotenoid actions on Th cells may vary in each 

carotenoid and do not seem to be closely associated with carotenoid antioxidant 

activity or relative polarity. 



Immunity 

201

Anticancer Res. 1999 Nov-Dec;19(6B):5223-7. 

Dietary beta-carotene and astaxanthin but not canthaxanthin stimulate 

splenocyte function in mice. 

Chew BP, Wong MW, Park JS, Wong TS. 

Department of Animal Sciences, Washington State University, Pullman 99164, 

USA. 

The in vivo modulatory effect of beta-carotene, astaxanthin and canthaxanthin on 

lymphocyte function was investigated. Female BALB/c mice (8 wk old) were fed 

a basal diet containing 0, 0.1% or 0.4% beta-carotene, astaxanthin or 

canthaxanthin for 0, 2 or 4 wk (n = 8/diet/period). Splenic lymphocytes were 

isolated and mitogen-stimulated proliferation, IL-2 production and lymphocyte 

cytotoxicity were assessed. Body weight and feed intake were not different among 

dietary treatments. Plasma carotenoids were undetectable in unsupplemented mice 

but concentrations of the respective carotenoids were elevated in mice fed 0.1 or 

0.4% beta-carotene (0.22 and 0.39 mumol/L), astaxanthin (16.4 and 50.2 

mumol/L) and canthaxanthin (5.00 and 7.02 mumol/L) respectively. Mice fed 

both dietary levels of beta-carotene and astaxanthin had enhanced 

phytohemagglutinin-induced lymphoblastogenesis compared to unsupplemented 

mice (P < 0.03). No treatment difference was detected with concanavalin A- or 

lipopolysaccharide-induced lympho-proliferation nor with IL-2 production (P < 

0.05). Astaxanthin (0.1%) also enhanced lymphocyte cytotoxic activity (P < 

0.08). In contrast, canthaxanthin did not significantly influence any of the 

lymphocyte functions measured. Results indicate that beta-carotene and 

astaxanthin but not canthaxanthin exert enhanced splenic lymphocyte function in 

mice. 

Immunity 

202

Br Poult Sci. 2007 Feb;48(1):90-7. 

Effect of dietary supplementation of astaxanthin by Phaffia 

rhodozyma on lipid peroxidation, drug metabolism and some 

immunological variables in male broiler chicks fed on diets with or 

without oxidised fat. 

Takimoto T, Takahashi K, Akiba Y. 

Laboratory of Animal Nutrition, Graduate School of Agricultural Science, 

Tohoku University, Sendai, Japan. 

1. Effects of dietary supplementation of astaxanthin (Ax) provided from Phaffia 

rhodozyma on lipid peroxidation, hepatic drug metabolism, antibody titres to 

sheep red blood cells (SRBC) and splenocyte proliferation to mitogens were 

determined in male broiler chicks. 2. Chicks, one week old, were given diets with 

or without oxidised fat (0 or 3.7 meq of peroxide value (POV)/kg diet) and/or Ax 

(0 or 100 mg/kg diet) for 14 d, ad libitum. 3. Lipid peroxidation, estimated by 2- 

thiobarbituric acid reactants values in liver, spleen, heart, plasma and hepatic 

microsomes, were increased by feeding a diet containing oxidised fat (P

Crit Rev Food Sci Nutr. 2006;46(2):185-96. 

Astaxanthin: a review of its chemistry and applications. 

Higuera-Ciapara I, Félix-Valenzuela L, Goycoolea FM. 

Centro de Investigación en Alimentación y Desarrollo, A.C., P.O. Box 1735. 

Hermosillo, Sonora, 83000, México. higuera@cascabel.ciad.mx 














recent and has mostly been performed in vitro or at the pre-clinical level with 

humans. This paper reviews the current available evidence regarding astaxanthin 

chemistry and its potential beneficial effects in humans. 



Immunity 

204

Fish Shellfish Immunol. 2004 Apr;16(4):527-37. 

Enhancement of innate immunity in rainbow trout (Oncorhynchus 

mykiss Walbaum) associated with dietary intake of carotenoids from 

natural products. 

Amar EC, Kiron V, Satoh S, Watanabe T. 

Tokyo University of Marine Science and Technology, Minato, Japan. 

The effects of orally administered carotenoids from natural sources on the nonspecific 

defense mechanisms of rainbow trout were evaluated in a nine-week 

feeding trial. Fish were fed four diets containing either beta-carotene or 

astaxanthin at 100 and 200 mg kg-1 from the marine algae Dunaliella salina and 

red yeast Phaffia rhodozyma, respectively, and a control diet containing no 

supplemented carotenoids. Specific growth rate and feed:gain ratio were not 

affected by dietary carotenoid supplementation. Among the humoral factors, 

serum alternative complement activity increased significantly in all carotenoid 

supplemented groups when compared to the control. On the other hand, serum 

lysozyme activity increased in the Dunaliella group but not in the Phaffia group, 

whereas plasma total immunoglobulin levels were not altered by the feeding 

treatments. As for the cellular responses, the superoxide anion production from 

the head kidney remained unchanged while the phagocytic rate and index in all 

supplemented groups were significantly higher than those of the control. These 

findings demonstrate that dietary carotenoids from both D. salina and P. 

rhodozyma can modulate some of the innate defense mechanisms in rainbow 

trout. 



Immunity 

205

J Nutr. 2004 Jan;134(1):257S-261S. 

Carotenoid action on the immune response. 

Chew BP, Park JS. 

Department of Animal Sciences, Washington State University, Pullman, WA 

99164-6351, USA. boonchew@wsu.edu 

Early studies demonstrating the ability of dietary carotenes to prevent infections 

have left open the possibility that the action of these carotenoids may be through 

their prior conversion to vitamin A. Subsequent studies to demonstrate the 

specific action of dietary carotenoids have used carotenoids without provitamin A 

activity such as lutein, canthaxanthin, lycopene and astaxanthin. In fact, these 

nonprovitamin A carotenoids were as active, and at times more active, than betacarotene 

in enhancing cell-mediated and humoral immune response in animals 

and humans. Another approach to study the possible specific role of dietary 

carotenoids has used animals that are inefficient converters of carotenoids to 

vitamin A, for example the domestic cat. Results have similarly shown immunoenhancement 

by nonprovitamin A carotenoids, based either on the relative 

activity or on the type of immune response affected compared to beta-carotene. 

Certain carotenoids, acting as antioxidants, can potentially reduce the toxic effects 

of reactive oxygen species (ROS). These ROS, and therefore carotenoids, have 

been implicated in the etiology of diseases such as cancer, cardiovascular and 

neurodegenerative diseases and aging. Recent studies on the role of carotenoids in 

gene regulation, apoptosis and angiogenesis have advanced our knowledge on the 

possible mechanism by which carotenoids regulate immune function and cancer. 



Immunity 

206

J Nutr. 1995 Oct;125(10):2483-92. 

Astaxanthin, a carotenoid without vitamin A activity, augments 

antibody responses in cultures including T-helper cell clones and 

suboptimal doses of antigen. 

Jyonouchi H, Sun S, Tomita Y, Gross MD. 


Minneapolis 55455, USA. 

Astaxanthin, a carotenoid without vitamin A activity, enhances T-dependent 

antigen (Ag)-specific humoral immune responses. We examined carotenoid 

actions on T-helper (Th) cell activity in a direct manner with reconstitution 

experiments; spleen Th cells were replaced with Ag-specific Type 1 and Type 2 

(Th1 and Th2) Th cell clones. The Ag for the Th1 and Th2 clones were pigeon 

cytochrome C and rabbit gamma-globulin, respectively. Astaxanthin and betacarotene 

augmented the number of IgM antibody (Ab)-secreting cells when 

unprimed B cells were incubated with Th clones and stimulated with suboptimal 

doses of Ag specific for each Th clone. The number of IgG Ab-secreting cells 

were greater with use of in vivo primed B cells than with unprimed B cells in both 

Th clones. Astaxanthin but not beta-carotene augmented the number of IgG Absecreting 

cells when primed B cells and Th cell clones were stimulated with 

suboptimal doses of Ag specific for each Th clone. In the presence of optimal 

doses of Ag for each Th clone, neither carotenoid augmented the number of Absecreting 

cells. Astaxanthin and beta-carotene may enhance the actions of both 

Th1 and Th2 cells for humoral immune responses with suboptimal Ag challenges; 

certain carotenoids may help maintain Ag-mediated immune responses at optimal 

levels. 



Immunity 

207

Nutr Cancer. 1995;23(2):171-83. 

Effect of carotenoids on in vitro immunoglobulin production by 

human peripheral blood mononuclear cells: astaxanthin, a 

carotenoid without vitamin A activity, enhances in vitro 

immunoglobulin production in response to a T-dependent stimulant 

and antigen. 

Jyonouchi H, Sun S, Gross M. 


Minneapolis 55455, USA. 

The effect of carotenoids on in vitro immunoglobulin (Ig) production by 

peripheral blood mononuclear cells (PBMNC) was examined by employing blood 

samples from adult volunteers and full-term newborn babies (umbilical cord 

blood). Under carotenoid-supplemented culture conditions, cells were stimulated 

by polyclonal stimulants, neoantigens, and a recall antigen (Ag), and IgM, IgA, 

and IgG levels in the culture supernatant were measured. Beta-carotene and 

astaxanthin were used as representatives of carotenoids with and without vitamin 

A activity, respectively. Astaxanthin enhanced IgM production in response to T- 

dependent Ag (TD-Ag) and a T-dependent polyclonal stimulant. Astaxanthin also 

augmented IgG production in response to a recall Ag. IgA production without 

supplemental carotenoids was negligible for all stimuli. However, in carotenoidsupplemented 

cultures, IgA production was significantly higher in response to a 

T-dependent polyclonal stimulant than in unsupplemented cultures. IgM and IgA 

production was augmented at 10(-8) mol/l astaxanthin, whereas astaxanthin 

enhanced IgG production in response to a recall Ag at 10(-10)-10(-9) mol/l. 

Similar enhancing actions of astaxanthin on IgM production were observed in 

cord blood mononuclear cells (CBMNC), although CBMNC produced less IgM 

than adult PBMNC. Beta-carotene did not have a significant effect on human Ig 

production. The carotenoid actions were not demonstrated under serum-free 

culture conditions; serum is essential for solubilization of carotenoids. In 

summary, this study has shown for the first time that astaxanthin, a carotenoid 

without vitamin A activity, enhances human Ig production in response to T- 

dependent stimuli. 


Immunity 

208

Nutr Cancer. 1994;21(1):47-58. 

Immunomodulating actions of carotenoids: enhancement of in vivo 

and in vitro antibody production to T-dependent antigens. 

Jyonouchi H, Zhang L, Gross M, Tomita Y. 

Department of Pediatrics, University of Minnesota, Minneapolis 55455. 

Previously, we demonstrated an enhancement of in vitro antibody (Ab) 

production in response to T-dependent antigens (TD-Ag) by astaxanthin, a 

carotenoid without vitamin A activity. The effects of beta-carotene, a carotenoid 

with vitamin A activity, and lutein, another carotenoid without vitamin A activity, 

on in vitro Ab production were examined with spleen cells from young and old 

B6 mice. In addition, the in vivo effects of lutein, astaxanthin, and beta-carotene 

on Ab production were studied in young and old B6 mice. Lutein, but not betacarotene, 

enhanced in vitro Ab production in response to TD-Ags. The depletion 

of T-helper cells prevented the enhancement of Ab production by lutein and 

astaxanthin. In vivo Ab production in response to TD-Ag was significantly 

enhanced by lutein, astaxanthin, and beta-carotene. The numbers of 

immunoglobulin M- and G-secreting cells also increased in vivo with the 

administration of these carotenoids when mice were primed with TD-Ags. 

Antibody production in response to TD-Ags in vivo and in vitro was significantly 

lower in old than in young B6 mice. Astaxanthin supplements partially restored 

decreased in vivo Ab production in response to TD-Ags in old B6 mice. Lutein 

and beta-carotene also enhanced in vivo Ab production in response to TD-Ags in 

old B6 mice, although to a lesser extent than did astaxanthin. However, none of 

the carotenoids had an effect on in vivo or in vitro Ab production in response to 

T-independent antigen. These results indicate significant immunomodulating 

actions of carotenoids for humoral immune responses to TD-Ags and suggest that 

carotenoid supplementation may be beneficial in restoring humoral immune 

responses in older animals. 



Immunity 

209

Nutr Cancer. 1993;19(3):269-80. 

Studies of immunomodulating actions of carotenoids. II. 

Astaxanthin enhances in vitro antibody production to T-dependent 

antigens without facilitating polyclonal B-cell activation. 

Jyonouchi H, Zhang L, Tomita Y. 

Department of Pediatrics, University of Minnesota, Minneapolis 55455. 

Previously we have shown that astaxanthin, a carotenoid without provitamin A 

activity, enhances in vitro antibody (Ab) production to sheep red blood cells in 

normal B6 mice. In this study, we further attempted to examine the mechanisms 

of this enhancing action of carotenoids on specific Ab production in vitro in 

relation to different antigen (Ag) stimuli, cytokine production, and T- and B-cell 

interactions in both normal and autoimmune strains of mice. When the actions of 

carotenoids were tested in normal strains of mice, we found that astaxanthin 

enhanced in vitro Ab production to T cell-dependent Ag, but not to T-independent 

Ag, and did not augment total immunoglobulin production. Astaxanthin exerted 

maximum enhancing actions when it was present at the initial period of Ag 

priming. This action of astaxanthin was abolished when T cells were depleted 

from spleen cell suspensions and appeared to require direct interactions between 

T and B cells. The results also indicated that carotenoids may modulate the 

production of interferon-tau in this assay system. When the actions of carotenoids 

were tested in autoimmune-prone MRL and NZB mice, the enhancing action of 

astaxanthin on in vitro Ab production was less significant. Furthermore, 

carotenoids did not potentiate or augment spontaneous Ab and immunoglobulin 

production by spleen cells in these strains. Taken together, carotenoids without 

provitamin A activity may be able to augment in vitro specific Ab production to T 

cell-dependent Ag partly through affecting the initial stage of Ag presentation 

without facilitating polyclonal B-cell activation or autoantibody production. 



Immunity 

210

Nutr Cancer. 1991;16(2):93-105. 

Studies of immunomodulating actions of carotenoids. I. Effects of 

beta-carotene and astaxanthin on murine lymphocyte functions and 

cell surface marker expression in in vitro culture system. 

Jyonouchi H, Hill RJ, Tomita Y, Good RA. 

Department of Pediatrics, University of South Florida/All Children's Hospital, St. 

Petersburg 33701. 

The immunomodulating effects of carotenoids (beta-carotene and astaxanthin) on 

mouse lymphocytes were studied in in vitro culture system by use of assay for 

mitogen responses of spleen cells, thymocyte proliferation, interleukin 2 

production, and antibody (Ab) production in vitro in response to sheep red blood 

cells. Changes of cell surface markers on spleen lymphocytes including Ia antigen 

(Ag), surface immunoglobulin, B220, and Thy-1 Ag were also examined. At a 

concentration of 10(-8) M, carotenoids did not show any significant effect on 

mitogen responses (phytohemagglutinin P and concanavalin A) on murine spleen 

cells, irrespective of the concentrations of mitogens used. Interleukin 2 production 

by murine spleen cells was not significantly altered by carotenoids in the culture 

media (10(-7) to 10(-9) M). [3H]thymidine incorporation by B6 thymocytes was 

somewhat enhanced in the presence of astaxanthin or beta-carotene when cultured 

in the concentration of 10(6)/ml. At higher concentrations of cells (5 x 10(6)/ml), 

such an effect was not observed. In assays of in vitro Ab production in response to 

sheep red blood cells, B6 spleen cells produced significantly more Ab-forming 

cells (plaque-forming cells, immunoglobulins M and G) in the presence of 

astaxanthin (greater than 10(-8) M) but not beta-carotene. Expression of Ia Ag 

seemed to be moderately enhanced on both Thy-1+ and Thy-1- spleen cells in the 

presence of astaxanthin (greater than 10(-9) M) but not beta-carotene. The 

expression of Thy-1 and surface immunoglobulin seemed unchanged with the 

treatment of these carotenoids. These results indicate that immunomodulating 

actions of carotenoids are not necessarily related to provitamin A activity, because 

astaxanthin, which does not have provitamin A activity, showed more significant 

effects in these bioassays and also indicate that such actions of carotenoid 

demonstrated in this study may be difficult to explain only by its oxygenquenching 

capacity. 



Immunity 

211

Clin Microbiol Infect. 2002 Jul;8(7):438-41. 

Effect of antioxidants on the immune response of Helicobacter 

pylori. 

Akyön Y. 

Hacettepe University, School of Medicine, Department of Microbiology and 

Clinical Microbiology, Ankara, Turkey. yakyon@tr.net 

Antioxidants are substances capable of inhibiting oxidation. In chronic diseases, 

inflammatory response cells produce oxygen free radicals. Oxygen free radicals 

cause DNA damage, and this may lead to gene modifications that might be 

carcinogenic. Chronic Helicobacter pylori infection causes the production of 

DNA-damaging free radicals. In recent years, various groups have studied the 

effects of antioxidants, especially on H. pylori-associated gastric cancer. In most 

of the studies, it has been shown that H. pylori infection does affect the level of 

antioxidants measured in the gastric juice, but there are also controversial results. 

Recent experimental studies, both in vivo and in vitro, have shown that vitamin C 

and astaxanthin, a carotenoid, are not only free radical scavengers but also show 

antimicrobial activity against H. pylori. It has been shown that astaxanthin 

changes the immune response to H. pylori by shifting the Th1 response towards a 

Th2 T-cell response. Very few experimental studies support the epidemiologic 

studies, and further studies are needed to describe the effect and the mechanism of 

antioxidants in the H. pylori immune response. 

Immunity 

212

NUTRITION AND CANCER, 36(1), 59-65 

. 

Antitumor Activity of Astaxanthin and Its Mode of Action 

Harumi Jyonouchl, Sinine Sun, KoJi [lJlma, and Myron D. Gross 

Astaxanthin, a carotenoid without Vitamin A activity, may excert antitumor 

activity through the enhancement of immune response. Here, we determined the 

effects of dietary astaxanthin on timor growth and tumor immunity against 

transplantable methylcholanthrene-induces fibrosarcoma (Meth-A tumor) cells. 

These tumor cells express a tumor antigen that induces T cell-mediated immune 

responses in syngenic mice. BALB/c mice were fed astaxanthin (0.02%, 40 

µg/kg body wt/day in a beadlet form) mixed in a chemically defined diet starting 

zero, one, and three weeks before subcutaneous inoculation with tumor cells (3 x 

10^5 cells, 2 times the minimal tumorigenic dose). Three weeks after inoculation, 

tumor size and weight were determined. We also determined cytotoic T 

lymphocyte (CTL) activity and interferon-γ (IFN-γ) production by tumor-draining 

lymph node (TDLN) and spleen cells by restimulating cells with Meth-A tumor 

cells in culture. The astaxanthin-fed mice had significantly lower tumor size and 

weight than controls when supplementation was started one and three weeks 

before tumor inoculation. This antitumor activity was paralleled with higher CTL 

activity and IFN-γ production by TNLN and spleen cells in the astaxanthin-fed 

mice. CTL activity by TDLN cells was highest in mice fed astaxanthin for three 

weeks before inoculation. When the astaxanthin-supplemented diet was started at 

the same time as tumor inoculation, none of these parameters were altered by 

dietary astaxanthin-supplemented diet was started at the same time as tumor 

inoculation, none of these parameters were altered by dietary astaxanthin, except 

IFN-γ production by spleen cells. Total serum astaxanthin concentrations were 

approximately 1.2 µmol/l when mice were fed astaxanthin (0.02%) for four weeks 

and appeared to increase in correlation with the length of astaxanthin 

supplementation. Our results indicate that dietary astaxanthin suppressed Meth-A 

tumor cell growth and stimulated immunity against Meth-A tumor antigen. 

Immunity 

213

J Clin Biochem Nutr. 2010 Sep;47(2):130-7. Epub 2010 Jun 22. 

Evaluation of therapeutic effects of astaxanthin on impairments in 

salivary secretion. 

Yamada T, Ryo K, Tai Y, Tamaki Y, Inoue H, Mishima K, Tsubota K, Saito I. 

Department of Pathology, Tsurumi University School of Dental Medicine, 2-1-3, 

Tsurumi, Tsurumi-ku, Yokohama 230-8501, Japan. 


The involvement of reactive oxygen species (ROS) in the pathophysiology of Sjögren's 

syndrome (SS), an autoimmune disorder, and irradiation-induced impairments in salivary 

secretion has been reported. Meanwhile, the strong antioxidant astaxanthin (Ast) has been 

suggested to have therapeutic effects on various diseases. In the present study, we 

examined the ROS scavenging capacity of Ast using a human salivary gland epithelial 

cell line (HSY) and investigated the effects of Ast on salivary secretion in a mouse model 

of irradiation-induced salivary gland dysfunction. Furthermore, we performed a clinical 

study of Ast in six SS patients and six normal individuals, quantifying the volume of 

saliva secretion and the level of oxidative stress markers in the saliva. Ast partially 

suppressed hydrogen peroxide-induced ROS in HSY cells. The mouse model 

demonstrated that the pre-administration of Ast resulted in the suppression of irradiationinduced 

hyposalivation. Furthermore, the administration of Ast appeared to increase 

salivary output in both the SS and normal groups. The level of oxidative stress marker, 

hexanoyl-lysine, in the saliva was reduced after Ast intake. These results suggest that Ast 

might act as an ROS scavenger, providing benefits to SS patients with impaired salivary 

secretion. 


Immunity 

214

Vet Immunol Immunopathol. 2011 Sep 3. [Epub ahead of print] 

Astaxanthin stimulates cell-mediated and humoral 

immune responses in cats. 

Park JS, Mathison BD, Hayek MG, Massimino S, Reinhart GA, Chew BP. 

Source 

School of Food Science, Washington State University, Pullman, WA 99164-6376, USA. 


Astaxanthin is a potent antioxidant carotenoid and may play a role in modulating immune 

response in cats. Blood was taken from female domestic shorthair cats (8-9mo old; 

3.2±0.04kg body weight) fed 0, 1, 5 or 10mg astaxanthin daily for 12wk to assess 

peripheral blood mononuclear cell (PBMC) proliferation response, leukocyte 

subpopulations, natural killer (NK) cell cytotoxic activity, and plasma IgG and IgM 

concentration. Cutaneous delayed-type hypersensitivity (DTH) response against 

concanavalin A and an attenuated polyvalent vaccine was assessed on wk 8 (prior to 

vaccination) and 12 (post-vaccination). There was a dose-related increase in plasma 

astaxanthin concentrations, with maximum concentrations observed on wk 12. Dietary 

astaxanthin enhanced DTH response to both the specific (vaccine) and nonspecific 

(concanavalin A) antigens. In addition, cats fed astaxanthin had heightened PBMC 

proliferation and NK cell cytotoxic activity. The population of CD3(+) total T and 

CD4(+) T helper cells were also higher in astaxanthin-fed cats; however, no treatment 

difference was found with the CD8(+) T cytotoxic and MHC II(+) activated lymphocyte 

cell populations. Dietary astaxanthin increased concentrations of plasma IgG and IgM. 

Therefore, dietary astaxanthin heightened cell-mediated and humoral immune responses 

in cats. 



Immunity 

215

Vet Immunol Immunopathol. 2011 Apr 15;140(3-4):199-206. Epub 2010 Dec 14. 

Dietary astaxanthin enhances immune response in dogs. 

Chew BP, Mathison BD, Hayek MG, Massimino S, Reinhart GA, Park JS. 

Source 

School of Food Science, Lewisburg, OH 45338, USA. boonchew@wsu.edu 


No information is available on the possible role of astaxanthin on immune response in 

domestic canine. Female Beagle dogs (9-10 mo old; 8.2 ± 0.2 kg body weight) were fed 

0, 10, 20 or 40 mg astaxanthin daily and blood sampled on wk 0, 6, 12, and 16 for 

assessing the following: lymphoproliferation, leukocyte subpopulations, natural killer 

(NK) cell cytotoxicity, and concentrations of blood astaxanthin, IgG, IgM and acute 

phase proteins. Delayed-type hypersensitivity (DTH) response was assessed on wk 0, 12 

and 16. Plasma astaxanthin increased dose-dependently and reached maximum 

concentrations on wk 6. Dietary astaxanthin enhanced DTH response to vaccine, 

concanavalin A-induced lymphocyte proliferation (with the 20mg dose at wk 12) and NK 

cell cytotoxic activity. In addition, dietary astaxanthin increased concentrations of IgG 

and IgM, and B cell population. Plasma concentrations of C reactive protein were lower 

in astaxanthin-fed dogs. Therefore, dietary astaxanthin heightened cell-mediated and 

humoral immune response and reduced DNA damage and inflammation in dogs. 



Immunity 

216

Cancer Prevention and Tumor Reduction 

Cancer Lett. 2009 May 5. [Epub ahead of print] 

Growth-inhibitory effects of the astaxanthin-rich alga 

Haematococcus pluvialis in human colon cancer cells. 

Palozza P, Torelli C, Boninsegna A, Simone R, Catalano A, Mele MC, Picci 

N. 

Institute of General Pathology, Catholic University School of Medicine, L. Go F. 

Vito, 1 00168 Rome, Italy. 

The growth-inhibitory effects of the astaxanthin-rich Haematococcus pluvialis 

were studied in HCT-116 colon cancer cells. H. pluvialis extract (5-25mug/ml) 

inhibited cell growth in a dose- and time-dependent manner, by arresting cell 

cycle progression and by promoting apoptosis. At 25mug/ml of H. pluvialis 

extract, an increase of p53, p21(WAF-1/CIP-1) and p27 expression (220%, 160%, 

250%, respectively) was observed, concomitantly with a decrease of cyclin D1 

expression (58%) and AKT phosphorylation (21%). Moreover, the extract, at the 

same concentration, strongly up-regulated apoptosis by modifying the ratio of 

Bax/Bcl-2 and Bcl-XL, and increased the phosphorylation of p38, JNK, and 

ERK1/2 by 160%, 242%, 280%, respectively. Growth-inhibitory effects by H. 

pluvialis were also observed in HT-29, LS-174, WiDr, SW-480 cells. This study 

suggests that H. pluvialis may protect from colon cancer. 


Cancer Prevention 

217

Toxicology. 2008 Jun 27;248(2-3):96-103. Epub 2008 Mar 27. 

Astaxanthin inhibits cytotoxic and genotoxic effects of 

cyclophosphamide in mice germ cells. 


Department of Pharmacology and Toxicology, National Institute of 

Pharmaceutical Education and Research, Sector-67, SAS. Nagar, Punjab 160062, 

India. 

Cyclophosphamide (CP), an alkylating agent used in the treatment of several 

cancers as well as an immunosuppressant in rheumatoid arthritis. It is used against 

several cancers due to its broad spectrum efficacy, but at the same time possesses 

unwanted risks for occupational exposure as well as therapy related toxicities to 

patients. The present study was aimed to investigate the protective effect of 

astaxanthin (AST) a red carotenoid pigment on CP induced germ cell toxicity in 

male mice. CP was administered intraperitoneally (i.p.) at the dose of 50, 100 and 

200mg/kg body weight to mice (20-25 g) once in a week for a period of five 

weeks. AST was given at the dose of 25mg/kg per oral (p.o.) for five consecutive 

days in a week for five weeks. The animals were sacrificed one week after the last 

injection of CP. The protective effect of AST against CP induced male germ cell 

toxicity was evaluated using body weight, testes and epididymis weight, sperm 

count, sperm head morphology, sperm comet assay, histology of testes and 

TUNEL assay. AST treatment significantly improved the testes weight, sperm 

count and sperm head morphology as compared to only CP treated animals. The 

result of comet assay showed that AST treatment significantly restored the sperm 

DNA damage induced by CP. Further, AST treatment showed protection against 

CP induced testicular toxicity as evident from testes histology and TUNEL assay. 

The present results indicate the chemoprotective potential of AST against CP 

induced germ cell toxicity in mice. 




218

Mol Nutr Food Res. 2006 Nov;50(11):991-5. 

Visualization of astaxanthin localization in HT29 human colon 

adenocarcinoma cells by combined confocal resonance Raman and 

fluorescence microspectroscopy. 

Briviba K, Bornemann R, Lemmer U. 

Lichttechnisches Institut and Center for Functional Nanostructures, Universität 

Karlsruhe TH, Karlsruhe, Germany. karlis.briviba@bfel.de 

Astaxanthin, a carotenoid found in plants and seafood, exhibits antiproliferative, 

antioxidant and anticarcinogenic properties. We show that astaxanthin delivered 

with tetrahydrofuran is effectively taken up by cultured colon adenocarcinoma 

cells and is localized mostly in the cytoplasm as detected by confocal resonance 

Raman and broad-band fluorescence microspectroscopy image analysis. Cells 

incubated with beta-carotene at the same concentration as astaxanthin (10 

microM) showed about a 50-fold lower cellular amount of beta-carotene, as 

detected by HPLC. No detectable Raman signal of beta-carotene was found in 

cells, but a weak broad-band fluorescence signal of beta-carotene was observed. 

beta-Carotene, like astaxanthin, was localized mostly in the cytoplasm. The 

heterogeneity of astaxanthin and beta-carotene cellular distribution in cells of 

intestinal origin suggests that the possible defense against reactive molecules by 

carotenoids in these cells may also be heterogeneous. 


 




219

Bioorg Med Chem. 2006 Aug 15;14(16):5451-8. Epub 2006 May 23. 

Molecular modeling of non-covalent binding of homochiral (3S,3'S)- 

astaxanthin to matrix metalloproteinase-13 (MMP-13). 

Bikádi Z, Hazai E, Zsila F, Lockwood SF. 

Virtua Drug, Ltd, Budapest, Hungary. 

Inhibitors for matrix metalloproteinases (MMPs) are under investigation for the 

treatment of various important chronic illnesses, including cancer, arthritis, and 

cardiovascular disease (CVD). In particular, MMP-13 is currently being probed as 

a potential key target in CVD and malignant disease due to its documented effects 

on extracellular matrix (ECM) remodeling, important in the pathophysiology of 

these diseases. Within the family of related mammalian MMP enzymes, MMP-13 

possesses a large hydrophobic binding pocket relative to that of other MMPs. 

Homochiral astaxanthin (3S,3'S-AST; 3S,3'S-dihydroxy-beta,beta-carotene-4,4'- 

dione), an important antioxidant and anti-inflammatory xanthophyll carotenoid, is 

an active metabolite of several novel soft drugs in clinical development; it is also 

extensively used and tested as a human nutraceutical. In the current study, the 

prediction of the geometry and energetics of its binding to human MMP-13 was 

conducted with molecular modeling. The method used was found to predict the 

energy of binding of known ligands of MMP-13 with great precision. Blind 

docking using the whole protein target was then used in order to identify the 

possible binding site(s) of AST. AST was predicted to bind at several sites in 

close proximity to the active center. Subsequent analyses focused on the binding 

site at the atomic (i.e., amino acid sequence) level suggested that AST can bind to 

MMP-13 with high affinity and favorable energetics. Therefore, the modeling 

study predicts potential direct enzyme-inhibitory activity of AST against MMP- 

13, a behavior that may be exploited in mammalian systems in which pathological 

upregulation of MMP activity is paramount. 



220

Int J Mol Med. 2005 Nov;16(5):931-6. 

Antiproliferation and induction of cell death of Phaffia rhodozyma 

(Xanthophyllomyces dendrorhous) extract fermented by brewer 

malt waste on breast cancer cells. 

Teo IT, Chui CH, Tang JC, Lau FY, Cheng GY, Wong RS, Kok SH, Cheng 

CH, Chan AS, Ho KP. 

State Key Laboratory of Chinese Medicine and Molecular Pharmacology, 

Shenzhen, Department of Applied Biology and Chemical Technology, The Hong 

Kong Polytechnic University, Hung Hom, Hong Kong, PR China. 

Astaxanthin has been shown to have antiproliferative activity on breast cancer and 

skin cancer cells. However, the high cost of production, isolation and purification 

of purified astaxanthin from natural sources or chemically synthetic methods limit 

its usage on cancer therapy. We show that astaxanthin could be produced by 

fermentating the Phaffia rhodozyma (Xanthophyllomyces dendrorhous) yeast 

cells with brewer malt waste using a 20 L B. Braun fermentor. The percentage 

composition of astaxanthin from the P. rhodozyma was >70% of total pigment as 

estimated by the high performance liquid chromatographic analysis. Furthermore, 

the antiproliferative activity of this P. rhodozyma cell extract (PRE) was 

demonstrated on breast cancer cell lines including the MCF-7 (estrogen receptor 

positive) and MDA-MB231 (estrogen receptor negative) by using the [3-(4,5- 

dimethylthiazol-2-yl)-5-(3-arboxymethoxyphenyl)-2- (4-sulfophenyl)-2Htetrazolium] 

(MTS) assay. No apoptotic cell death, but growth inhibitory effect 

was induced after 48 h of PRE incubation as suggested by morphological 

investigation. Anchorage-dependent clonogenicity assay showed that PRE could 

reduce the colony formation potential of both breast cancer cell lines. Cell death 

was observed from both breast cancer cell lines after incubation with PRE for 6 

days. Taken together, our results showed that by using an economic method of 

brewer malt waste fermentation, we obtained P. rhodozyma with a high yield of 

astaxanthin and the corresponding PRE could have short-term growth inhibition 

and long-term cell death activity on breast cancer cells. 




221

Biochim Biophys Acta. 2005 May 30;1740(2):170-8. Epub 2005 Jan 25. 

Cancer prevention by retinoids and carotenoids: independent action 

on a common target. 

Bertram JS, Vine AL. 

Cancer Research Center of Hawaii, University of Hawaii at Manoa, 1236 Lauhala 

St., Honolulu, Hawaii, USA. john@crch.hawaii.edu 

Virtually all human tumors are deficient in gap junctional communication (GJC) 

and the restoration of GJC by forced expression of connexins reduces indices of 

neoplasia. The expression of connexin 43 (Cx43) is upregulated by cancerpreventive 

retinoids and carotenoids which correlates with the suppression of 

carcinogen-induced transformation in 10T1/2 cells. However, the molecular 

mechanism for upregulated expression is poorly understood. The retinoic acid 

receptor antagonist, Ro 41-5253, suppressed retinoid-induced Cx43 protein 

expression in 10T1/2 cells and the induction of a Cx43 luciferase reporter 

construct in F9 cells, but did not suppress protein expression or reporter activity 

induced by the non-pro-vitamin A carotenoid astaxanthin. In contrast, Cx43 

induction by astaxanthin, but not by a RAR-specific retinoid, was inhibited by 

GW9662, a PPAR-gamma antagonist. Neither compound required protein 

synthesis for the induction of Cx43 mRNA, nor was the 5.0 h half-life of Cx43 

mRNA altered, indicating direct transcriptional activation. The responsive region 

was found within -158 bp and +209 bp of the transcription start site. Site directed 

mutagenesis of a GC-box in this region increased basal levels of transcription and 

loss of retinoid responsiveness. Simultaneous treatment with a retinoid and betacarotene 

or astaxanthin resulted in supra-additive Cx43 expression, again 

indicating separate mechanisms of gene regulation. 




222

Carcinogenesis. 2005 Sep;26(9):1634-41. Epub 2005 May 11. 

Inhibition of chemically-induced neoplastic transformation by a 

novel tetrasodium diphosphate astaxanthin derivative. 

Hix LM, Frey DA, McLaws MD, Østerlie M, Lockwood SF, Bertram JS. 

Department of Cell and Molecular Biology, University of Hawaii at Manoa, 

Honolulu, HI 96822, USA. 

Carotenoids have been implicated in numerous epidemiological studies as being 

protective against cancer at many sites, and their chemopreventive properties have 

been confirmed in laboratory studies. Astaxanthin (AST), primarily a carotenoid 

of marine origin, responsible for the pink coloration of salmon, shrimp and 

lobster, has received relatively little attention. As with other carotenoids, its 

highly lipophilic properties complicate delivery to model systems. To overcome 

this issue we have synthesized a novel tetrasodium diphosphate astaxanthin 

(pAST) derivative with aqueous dispersibility of 25.21 mg/ml. pAST was 

delivered to C3H/10T1/2 cells in an aqueous/ethanol solution and compared with 

non-esterified AST dissolved in tetrahydrofuran. We show pAST to (i) upregulate 

connexin 43 (Cx43) protein expression; (ii) increase the formation of Cx43 

immunoreactive plaques; (iii) upregulate gap junctional intercellular 

communication (GJIC); and (iv) cause 100% inhibition of methylcholanthreneinduced 

neoplastic transformation at 10(-6) M. In all these assays, pAST was 

superior to non-esterified AST itself; in fact, pAST exceeded the potency of all 

other previously tested carotenoids in this model system. Cleavage of pAST to 

non-esterified (free) AST and uptake into cells was also verified by HPLC; 

however, levels of free AST were approximately 100-fold lower than in cells 

treated with AST itself, suggesting that pAST possesses intrinsic activity. The 

dual properties of water dispersibility (enabling parenteral administration in vivo) 

and increased potency should prove extremely useful in the future development of 

cancer chemopreventive agents. 



223

Cancer Lett. 2004 Jul 28;211(1):25-37. 

Upregulation of connexin 43 protein expression and increased gap 

junctional communication by water soluble disodium disuccinate 

astaxanthin derivatives. 

Hix LM, Lockwood SF, Bertram JS. 

Department of Cell and Molecular Biology, University of Hawaii at Manoa, 

Honolulu 96822, USA. 

Carotenoids are plant pigments whose consumption is associated with lower 

cancer rates in humans. Studies in experimental animal and cell systems have 

confirmed the cancer chemopreventive activity of these compounds. However, 

their extremely hydrophobic nature makes these compounds biologically 

unavailable unless delivered in organic solution to model systems. We have 

synthesized novel disodium salt disuccinate astaxanthin derivatives that possess 

high aqueous dispersibility. When delivered to mouse embryonic fibroblast 

C3H/10T1/2 cell cultures, either in aqueous or aqueous/ethanol solutions, these 

derivatives are biologically active. Biological activity was demonstrated by (1) 

upregulated expression of connexin 43 (Cx43) protein; (2) increased formation of 

Cx43 immunoreactive plaques in regions of the plasma membrane consistent with 

localization of gap junctions; (3) significantly upregulated gap junctional 

intercellular communication (GJIC) as demonstrated by Lucifer Yellow dye 

transfer after microinjection (P < 0.03; Fisher's Exact test). Enhanced expression 

of Cx43 and increased GJIC have been previously demonstrated to result in 

inhibition of in vitro neoplastic transformation of 10T1/2 cells as well as growth 

reduction of human tumors in xenografts. These novel derivatives possess 

increased utility as water soluble and water dispersible agents, allowing for 

aqueous delivery both in vitro and in vivo, properties that could enhance their 

potential clinical utility as potent cancer chemopreventive agents. Copyright 2004 

Elsevier Ireland Ltd. 



224

Life Sci. 2002 Apr 21;70(21):2509-20. 

Contribution of the antioxidative property of astaxanthin to its 

protective effect on the promotion of cancer metastasis in mice 

treated with restraint stress. 

Kurihara H, Koda H, Asami S, Kiso Y, Tanaka T. 

Institute for Health Care Science, Suntory Ltd., 1-1-1 Wakayamadai, Shimamotocho, 

Mishima-gun, Osaka 618-8503, Japan. Hiroshi_Kurihara@suntory.co.jp 

We investigated the effects of astaxanthin on the antitumor effector activity of 

natural killer (NK) cells suppressed by stress in mice in order to define the 

immunological significance of astaxanthin (ASX) when combined with restraint 

stress treatment. When the mice were treated with restraint stress alone, the total 

number of spleen cells, and the level NK cell activity per spleen were reduced to a 

nadir on day 3. The stress also caused a significant increase in the lipid 

peroxidation of liver tissue. ASX (100 mg/kg/day, p.o., 4 days) improved the 

immunological dysfunction induced by restraint stress. On the other hand, 

metastatic nodules were observed in the livers of syngenic DBA/2 mice on day 12 

after inoculation of P815 mastocytoma cells. Hepatic metastasis was promoted 

further by restraint stress when applied on day 3 before the inoculation of P815. 

Daily oral administration of ASX (1 mg/kg/day, p.o., 14 days) markedly 

attenuated the promotion of hepatic metastasis induced by restraint stress. These 

results suggested that astaxanthin improves antitumor immune responses by 

inhibiting of lipid peroxidation induced by stress. 



225

Nutr Cancer. 2000;36(1):59-65. 

Antitumor activity of astaxanthin and its mode of action. 

Jyonouchi H, Sun S, Iijima K, Gross MD. 


Minneapolis 55455, USA. jyono001@jyono001.email.umn.edu 

Astaxanthin, a carotenoid without vitamin A activity, may exert antitumor activity 

through the enhancement of immune responses. Here, we determined the effects 

of dietary astaxanthin on tumor growth and tumor immunity against 

transplantable methylcholanthrene-induced fibrosarcoma (Meth-A tumor) cells. 

These tumor cells express a tumor antigen that induces T cell-mediated immune 

responses in syngenic mice. BALB/c mice were fed astaxanthin (0.02%, 40 

micrograms/kg body wt/day in a beadlet form) mixed in a chemically defined diet 

starting zero, one, and three weeks before subcutaneous inoculation with tumor 

cells (3 x 10(5) cells, 2 times the minimal tumorigenic dose). Three weeks after 

inoculation, tumor size and weight were determined. We also determined 

cytotoxic T lymphocyte (CTL) activity and interferon-gamma (IFN-gamma) 

production by tumor-draining lymph node (TDLN) and spleen cells by 

restimulating cells with Meth-A tumor cells in culture. The astaxanthin-fed mice 

had significantly lower tumor size and weight than controls when 

supplementation was started one and three weeks before tumor inoculation. This 

antitumor activity was paralleled with higher CTL activity and IFN-gamma 

production by TDLN and spleen cells in the astaxanthin-fed mice. CTL activity 

by TDLN cells was highest in mice fed astaxanthin for three weeks before 

inoculation. When the astaxanthin-supplemented diet was started at the same time 

as tumor inoculation, none of these parameters were altered by dietary 

astaxanthin, except IFN-gamma production by spleen cells. Total serum 

astaxanthin concentrations were approximately 1.2 mumol/l when mice were fed 

astaxanthin (0.02%) for four weeks and appeared to increase in correlation with 

the length of astaxanthin supplementation. Our results indicate that dietary 

astaxanthin suppressed Meth-A tumor cell growth and stimulated immunity 

against Meth-A tumor antigen. 




226

Cancer Lett. 2000 Apr 3;151(1):111-5. 

Inhibitory effects of carotenoids on the invasion of rat ascites 

hepatoma cells in culture. 

Kozuki Y, Miura Y, Yagasaki K. 

Department of Applied Biological Science, Tokyo Noko University, Fuchu, 

Japan. 

The effects of carotenoids--alpha-carotene, beta-carotene, lycopene, betacryptoxanthin, 

zeaxanthin, lutein, canthaxanthin, astaxanthin--on the invasion of 

rat ascites hepatoma AH109A cells were investigated by co-culturing the 

hepatoma cells with rat mesentery-derived mesothelial cells (M-cells). All the 

carotenoids examined inhibited AH109A invasion in a dose-dependent manner up 

to 5 microM. Cancer cells previously cultured with hypoxanthine (HX) and 

xanthine oxidase (XO) showed a highly invasive activity. Carotenoids, 5 microM 

of beta-carotene and astaxanthin, suppressed this reactive oxygen speciespotentiated 

invasive capacity by simultaneously treating AH109A cells with the 

carotenoids, HX and XO. These results suggest that the antioxidative property of 

these carotenoids may be involved in their anti-invasive action. 




227

Anticancer Res. 1999 Nov-Dec;19(6B):5223-7. 

Dietary beta-carotene and astaxanthin but not canthaxanthin 

stimulate splenocyte function in mice. 

Chew BP, Wong MW, Park JS, Wong TS. 

Department of Animal Sciences, Washington State University, Pullman 99164, 

USA. 

The in vivo modulatory effect of beta-carotene, astaxanthin and canthaxanthin on 

lymphocyte function was investigated. Female BALB/c mice (8 wk old) were fed 

a basal diet containing 0, 0.1% or 0.4% beta-carotene, astaxanthin or 

canthaxanthin for 0, 2 or 4 wk (n = 8/diet/period). Splenic lymphocytes were 

isolated and mitogen-stimulated proliferation, IL-2 production and lymphocyte 

cytotoxicity were assessed. Body weight and feed intake were not different among 

dietary treatments. Plasma carotenoids were undetectable in unsupplemented mice 

but concentrations of the respective carotenoids were elevated in mice fed 0.1 or 

0.4% beta-carotene (0.22 and 0.39 mumol/L), astaxanthin (16.4 and 50.2 

mumol/L) and canthaxanthin (5.00 and 7.02 mumol/L) respectively. Mice fed 

both dietary levels of beta-carotene and astaxanthin had enhanced 

phytohemagglutinin-induced lymphoblastogenesis compared to unsupplemented 

mice (P < 0.03). No treatment difference was detected with concanavalin A- or 

lipopolysaccharide-induced lympho-proliferation nor with IL-2 production (P < 

0.05). Astaxanthin (0.1%) also enhanced lymphocyte cytotoxic activity (P < 

0.08). In contrast, canthaxanthin did not significantly influence any of the 

lymphocyte functions measured. Results indicate that beta-carotene and 

astaxanthin but not canthaxanthin exert enhanced splenic lymphocyte function in 

mice. 




228

Anticancer Res. 1999 May-Jun;19(3A):1849-53. 

A comparison of the anticancer activities of dietary beta-carotene, 

canthaxanthin and astaxanthin in mice in vivo. 

Chew BP, Park JS, Wong MW, Wong TS. 

Department Animal Sciences, Washington State University, Pullman 99164-6320, 

USA. boonchew@wsu.edu 

The anticancer activities of beta-carotene, astaxanthin and canthaxanthin against 

the growth of mammary tumors were studied in female eight-wk-old BALB/c 

mice. The mice were fed a synthetic diet containing 0, 0.1 or 0.4% beta-carotene, 

astaxanthin or canthaxanthin. After 3 weeks, all mice were inoculated with 1 x 

10(6) WAZ-2T tumor cells into the mammary fat pad. All animals were killed on 

45 d after inoculation with the tumor cells. No carotenoids were detectable in the 

plasma or tumor tissues of unsupplemented mice. Concentrations of plasma 

astaxanthin (20 to 28 mumol/L) were greater (P < 0.05) than that of beta-carotene 

(0.1 to 0.2 mumol/L) and canthaxanthin (3 to 6 mmol/L). However, in tumor 

tissues, the concentration of canthaxanthin (4.9 to 6.0 nmol/g) was higher than 

that of beta-carotene (0.2 to 0.5 nmol/g) and astaxanthin (1.2 to 2.7 nmol/g). In 

general, all three carotenoids decreased mammary tumor volume. Mammary 

tumor growth inhibition by astaxanthin was dose-dependent and was higher than 

that of canthaxanthin and beta-carotene. Mice fed 0.4% beta-carotene or 

canthaxanthin did not show further increases in tumor growth inhibition compared 

to those fed 0.1% of each carotenoid. Lipid peroxidation activity in tumors was 

lower (P < 0.05) in mice fed 0.4% astaxanthin, but not in those fed beta-carotene 

and canthaxanthin. Therefore, beta-carotene, canthaxanthin and especially 

astaxanthin inhibit the growth of mammary tumors in mice; their anti-tumor 

activity is also influenced by the supplemental dose. 




229

Br J Nutr. 1999 Mar;81(3):235-42. 

Effect of dietary supplementation with carotenoids on xenobiotic 

metabolizing enzymes in the liver, lung, kidney and small intestine 

of the rat. 

Jewell C, O'Brien NM. 

Department of Nutrition, University College, Cork, Ireland. 

The effect of 16 d intake of 300 mg carotenoids/kg diet (beta-carotene (beta C), 

bixin (BX), lycopene (LY), lutein (LU), canthaxanthin (CX) or astaxanthin (AX) 

on xenobiotic metabolizing enzymes in the liver, lung, kidney and small intestine 

of male Wistar rats was assessed. A control group received the basal diet (AIN- 

76) without carotenoids and a positive control group for enzyme induction 

received 3-methylcholanthrene (3-MC) at 666 mg/kg diet. Cytochrome P450 

activity was assessed using the substrates ethoxyresorufin for P450 1A1, 

methoxyresorufin for P450 1A2, pentoxyresorufin for P450 2B1/2 and 

benzyloxyresorufin for P450 types 1A1/2, 2B1/2 and 3A. Glutathione-Stransferase 

(EC 2.5.1.18) and reduced glutathione status were assessed. 

Carotenoid uptake by the tissues was also determined. 3-MC and the carotenoids 

BX, CX and AX led to significant increases compared with control in liver, lung 

and kidney ethoxyresorufin-O-deethylation. Methoxyresorufin-O-demethylation 

activity was significantly increased in liver and lung by BX, CX and AX but only 

CX and AX significantly increased activity in kidney. Pentoxyresorufin-Odepentylation 

and benzyloxyresorufin-O-dearylation increased in liver of 3-MC-, 

BX-, CX- and AX-treated rats, but to a much lesser degree than for the other two 

substrates. Benzyloxyresorufin-O-dearylation in lung was significantly decreased 

by all carotenoids. Activities of any of the measured enzymes in the small 

intestine were undetectable in all treatment groups except the 3-MC group. 

Glutathione status was unaffected by any of the treatments. This is the first study 

identifying the carotenoids BX, CX and AX as inducers of rat lung and kidney 

xenobiotic metabolizing enzymes. 




230

Carcinogenesis. 1998 Mar;19(3):403-11. 

Dietary carotenoids inhibit aflatoxin B1-induced liver preneoplastic 

foci and DNA damage in the rat: role of the modulation of aflatoxin 

B1 metabolism. 

Gradelet S, Le Bon AM, Bergès R, Suschetet M, Astorg P. 

Institut National de la Recherche Agronomique, Unité de Toxicologie 

Nutritionelle, Dijon, France. 

To study the effects of carotenoids on the initiation of liver carcinogenesis by 

aflatoxin B1 (AFB1), male weanling rats were fed beta-carotene, beta-apo-8'- 

carotenal, canthaxanthin, astaxanthin or lycopene (300 mg/kg diet), or an excess 

of vitamin A (21000 RE/kg diet), or were injected i.p. with 3-methylcholanthrene 

(3-MC) (6 x 20 mg/kg body wt) before and during i.p. treatment with AFB1 (2 x 1 

mg/kg body wt). The rats were later submitted to 2-acetylaminofluorene treatment 

and partial hepatectomy, and placental glutathione S-transferase-positive liver 

foci were detected and quantified. The in vivo effects of carotenoids or of 3-MC 

on AFB1-induced liver DNA damage were evaluated using different endpoints: 

liver DNA single-strand breaks (SSB) induced by AFB1, and in vivo binding of 

[3H]AFB1 to liver DNA and plasma albumin. Finally, the modulation of AFB1 

metabolism by carotenoids or by 3-MC was investigated in vitro by incubating 

[14C]AFB1 with liver microsomes from rats that had been fed with carotenoids or 

treated by 3-MC, and the metabolites formed by HPLC were analyzed. In contrast 

to lycopene or to an excess of vitamin A, both of which had no effect, betacarotene, 

beta-apo-8'carotenal, astaxanthin and canthaxanthin, as well as 3-MC, 

were very efficient in reducing the number and the size of liver preneoplastic foci. 

In a similar way as 3-MC, the P4501A-inducer carotenoids, beta-apo-8'-carotenal 

astaxanthin and canthaxanthin, decreased in vivo AFB1-induced DNA SSB and 

the binding of AFB1 to liver DNA and plasma albumin, and increased in vitro 

AFB1 metabolism to aflatoxin M1, a less genotoxic metabolite. It is concluded 

that these carotenoids exert their protective effect through the deviation of AFB1 

metabolism towards detoxication pathways. In contrast, beta-carotene did not 

protect hepatic DNA from AFB1-induced alterations, and caused only minor 

changes of AFB1 metabolism: seemingly, its protective effect against the 

initiation of liver preneoplastic foci by AFB1 is mediated by other mechanisms. 




231

Cancer Lett. 1997 Mar 19;114(1-2):221-3. 

Modulation of aflatoxin B1 carcinogenicity, genotoxicity and 

metabolism in rat liver by dietary carotenoids: evidence for a 

protective effect of CYP1A inducers. 

Gradelet S, Astorg P, Le Bon AM, Bergès R, Suschetet M. 

Unité de Toxicologie Nutritionnelle, INRA, Dijon, France. 

The effects of several carotenoids of vitamin A and of 3-methylcholanthrene have 

been tested on the initiation of hepatocarcinogenesis by aflatoxin B1, using the 

sequential protocol of Solt and Farber. AFB1-induced DNA single-strand breaks 

and AFB1-metabolism were also assessed. The P4501A inducer carotenoids 

(canthaxanthin, astaxanthin, beta-apo-8'-carotenal) and 3-methylcholanthrene 

reduce the carcinogenicity of AFB1, divert AFB1-metabolism into the less 

genotoxic aflatoxin M1 and reduce AFB1-induced DNA single-strand breaks: we 

conclude that these carotenoids exert their protective effect through the deviation 

of AFB1 metabolism towards detoxification pathways. beta-Carotene decreased 

AFB1 carcinogenicity but did not alter its metabolism, probably acting by other 

mechanisms. 




232

J Cell Biochem Suppl. 1997;27:35-41. 

Chemoprevention by naturally occurring and synthetic agents in 

oral, liver, and large bowel carcinogenesis. 

Mori H, Tanaka T, Sugie S, Yoshimi N, Kawamori T, Hirose Y, Ohnishi M. 

Department of Pathology, Gifu University School of Medicine, Japan. 

A number of naturally occurring compounds and several related synthetic agents 

were confirmed to exert chemopreventive properties against carcinogenesis in the 

digestive organs. Phenolic compounds, widely distributed as plant constituents, 

possess chemopreventive activities in tongue, liver, and large bowel of rodents. 

Of them, a simple phenolic protocatechuic acid seems to be a promising 

compound. Organosulfur compounds contained in the cruciferous vegetables and 

known to activate detoxifying enzymes are regarded as a candidate group for 

cancer preventive agents. We proved a strong protective effect of S- 

methylmethanethiosulfonate, a constituent in these vegetables, on azoxymethane 

(AOM)-induced large bowel carcinogenesis. Some oxygenated carotenoids 

(xanthophylls) are reported to have antitumor effects. Naturally occurring 

xanthophylls astaxanthin and canthaxanthin have considerable preventive 

activities on 4-nitroquinoline-1-oxide (4-NQO)-induced tongue carcinogenesis 

and AOM-induced large bowel carcinogenesis. A novel synthesized retinoidal 

butenolide, KYN-54, which suppresses large bowel as well as tongue 

carcinogenesis could be a useful agent for prevention of digestive organ cancers. 

Some trace elements are known to have anticarcinogenic effects. Magnesium 

hydroxide, a protective agent in colorectal carcinogenesis, inhibits c-myc 

expression and ornithine decarboxylase activity in the mucosal epithelium of the 

intestine. Our results show that many agents with preventive effects in tongue, 

liver, and large bowel control carcinogen-induced hyperproliferation of cells in 

these organs. Carcinogens used to induce large bowel cancers also induce 

apoptosis in the target sites. Telomerase activity is increased in the tissues of 

preneoplastic as well as neoplastic lesions in experimental models such as 

dimethylbenz[a]anthracene-induced oral carcinogenesis in hamsters. These could 

be useful biomarkers in studies for cancer chemoprevention. 




233

Carcinogenesis. 1995 Dec;16(12):2957-63. 

Suppression of azoxymethane-induced rat colon carcinogenesis by 

dietary administration of naturally occurring xanthophylls 

astaxanthin and canthaxanthin during the postinitiation phase. 

Tanaka T, Kawamori T, Ohnishi M, Makita H, Mori H, Satoh K, Hara A. 

First Department of Pathology, Gifu University School of Medicine, Japan. 

The modulating effects of dietary feeding of two xanthophylls, astaxanthin (AX) 

and canthaxanthin (CX) during the postinitiation phase on colon carcinogenesis 

initiated with azoxymethane (AOM) were investigated in male F344 rats. Animals 

were initiated with AOM by weekly s.c. injections of 15 mg/kg body wt for 3 

weeks and then they were fed the diets containing AX or CX at concentrations of 

100 and 500 p.p.m. for 34 weeks. The others contained the groups of rats treated 

with AX or CX alone and untreated. At the end of the study (week 37), the 

incidence and multiplicity of neoplasms (adenoma and adenocarcinoma) in the 

large intestine of rats initiated with AOM and followed by AX or CX containing 

diet at a high dose (500 p.p.m.) were significantly smaller than those of rats given 

AOM alone (P < 0.001). In addition, AX or CX feeding significantly inhibited the 

development of aberrant crypt foci induced by AOM. Dietary exposure to AX or 

CX also decreased cell proliferation activity as revealed by measuring 5'- 

bromodeoxyuridine-labeling index as crypt cells, colonic mucosal ornithine 

decarboxylase activity and blood polyamine levels. These results indicate that AX 

and CX are possible chemopreventers for carcinogenesis of colon in addition to 

urinary bladder and oral cavity and such effects may be partly due to suppression 

of cell proliferation. 




234

Cancer Res. 1995 Sep 15;55(18):4059-64. 

Chemoprevention of rat oral carcinogenesis by naturally occurring 

xanthophylls, astaxanthin and canthaxanthin. 

Tanaka T, Makita H, Ohnishi M, Mori H, Satoh K, Hara A. 


The chemopreventive effects of two xanthophylls, astaxanthin (AX) and 

canthaxanthin (CX) on oral carcinogenesis induced by 4-nitroquinoline 1-oxide 

(4-NQO) was investigated in male F344 rats. Rats were given 20 ppm of 4-NQO 

in their drinking water for 8 weeks to induce oral neoplasms or preneoplasms. 

Animals were fed diets containing 100 ppm AX or CX during the initiation or 

postinitiation phase of 4-NQO-induced oral carcinogenesis. The others contained 

the groups of rats treated with AX or CX alone and untreated. At the end of the 

study (week 32), the incidences of preneoplastic lesions and neoplasms in the oral 

cavity of rats treated with 4-NQO and AX or CX were significantly smaller than 

those of rats given 4-NQO alone (P < 0.001). In particular, no oral neoplasms 

developed in rats fed AX and CX during the 4-NQO exposure and in those given 

CX after the 4-NQO administration. Similarly, the incidences of oral 

preneoplastic lesions (hyperplasia and dysplasia) in rats treated with 4-NQO and 

AX or CX were significantly smaller than that of the 4-NQO-alone group (P < 

0.05). In addition to such tumor inhibitory potential, dietary exposure of AX or 

CX decreased cell proliferation activity in the nonlesional squamous epithelium 

exposed to 4-NQO as revealed by measuring the silver-stained nucleolar 

organizer regions protein number/nucleus and 5'-bromodeoxyuridine-labeling 

index. Also, dietary AX and CX could reduce polyamine levels of oral mucosal 

tissues exposed to 4-NQO. These results indicate that AX and CX are possible 

chemopreventers for oral carcinogenesis, and such effects may be partly due to 

suppression of cell proliferation. 




235

Carcinogenesis. 1994 Jan;15(1):15-9. 

Chemoprevention of mouse urinary bladder carcinogenesis by the 

naturally occurring carotenoid astaxanthin. 

Tanaka T, Morishita Y, Suzui M, Kojima T, Okumura A, Mori H. 


The chemopreventive effects of two xanthophylls, astaxanthin (AX) and 

canthaxanthin (CX), on urinary bladder carcinogenesis induced by N-butyl-N(4- 

hydroxybutyl)nitrosamine (OH-BBN) was investigated in male ICR mice. Mice 

were given 250 p.p.m. OH-BBN in drinking water for 20 weeks and after a 1 

week interval with tap water, water containing AX or CX at a concentration of 50 

p.p.m. was administered during subsequent 20 weeks. Other groups of mice were 

treated with AX or CX alone or untreated. At the end of the study (week 41), the 

incidences of preneoplastic lesions and neoplasms in the bladder of mice treated 

with OH-BBN and AX or CX were smaller than those of mice given OH-BBN. In 

particular, AX administration after OH-BBN exposure significantly reduced the 

incidence of bladder cancer (transitional cell carcinoma) (P < 0.003). However, 

the inhibition of the frequencies of such lesions in mice treated with OH-BBN and 

CX was not significant. Treatment with AX or CX also decreased the 

number/nucleus of silver-stained nucleolar organizer region proteins (AgNORs), a 

new index of cell proliferation, in the transitional epithelium exposed to OH- 

BBN. Preneoplasms and neoplasms induced by OH-BBN, and the 

antiproliferative potential, was greater for AX than CX. These results indicate that 

AX is a possible chemopreventive agent for bladder carcinogenesis and such an 

effect of AX may be partly due to suppression of cell proliferation. 




236

Autoimmunity. 1993;16(2):95-102. 

Preventive action of carotenoids on the development of 

lymphadenopathy and proteinuria in MRL-lpr/lpr mice. 

Tomita Y, Jyonouchi H, Engelman RW, Day NK, Good RA. 

Department of Public Health, School of Medicine, Kurume University, Japan. 

The chemopreventive action of carotenoids on proteinuria and lymphadenopathy 

were examined in autoimmune-prone MRL-lpr/lpr (MRL/l) mice. They were fed 

a synthetic full-fed diet (16-18 kcal/mouse/day) with supplementation of betacarotene 

or astaxanthin (0.19 mumoles/mouse, 3 times a week), and the 

development of lymphadenopathy and proteinuria were examined. MRL/l mice 

fed a full-fed diet without the supplementation of carotenoids or those fed a 

calorie-restricted (CR) diet (10-11 kcal/mouse/day, 60% calorie intake of full-fed 

mice) were employed as controls. CR dramatically delayed the development of 

proteinuria and lymphadenopathy, as reported previously. Carotenoids also 

significantly delayed the onset of these symptoms in MRL/l mice fed a full-fed 

diet. Carotenoids were half as effective as CR and astaxanthin, a carotenoid 

without provitamin A activity, which appeared to exert more significant 

preventive actions than beta-carotene in delaying the development of these 

symptoms. Similar chemopreventive actions of carotenoids were also 

demonstrated in MRL/l mice fed a regular diet (Lab Chow). CR has been shown 

to augment IL-2 production and to decrease serum prolactin levels in this strain, 

which may be related to its dramatic preventive action of autoimmunity. 

However, carotenoids did not affect IL-2 production nor prolactin levels in fullfed 

MRL/l mice. The chemopreventive actions of carotenoids observed in 

autoimmune-prone MRL/l mice may be attributed to yet unknown mechanisms, 

apart from their provitamin A activity or oxygen-quenching activity. 




237

Agricultural Chemistry & Biotechnology. 40(6): 490-494. #17. Lee, S. et al. (1997). 

Inhibition of benzo(a)pyrene-induced mouse forestomach neoplasia 

by astaxanthin containing egg yolks 

Anticarcinogenic activity of astaxanthin-containing egg yolks (designate AEY) 

was investigated for benzo(a)pyrene (BP)-induced mouse forestomach 

tumorigenesis initiating regimen. Female ICR mouse (6-7 weeks of age) were 

housed in polycarbonated cages (5 mice/cage; 20 mice/treatment) in a humidityand-temperature-controlled 

facility and permitted free access to water and food. 

One week later, four and 2 days prior to p.o. treatment with BP (2 mg/0.2 ml corn 

oil), mice were given 0.2 ml PBS containing 50 mg AEY, 100 mg AEY, 150 mg 

AEY, or 150 mg CEY. Control mice were only given 0.2 ml PBS. Three days 

later this sequence was repeated for a total of 4 times. Beginning with the first 

intubation and continuing thereafter, body weight and food intake were recorded 

once weekly. All surviving mice were sacrificed 24 weeks after the first dose of 

BP. Mice treated with AEY developed only about one third as many 

neoplasms/animal as mice in control or CEY-treated group (p

Journal of Kyoto Prefectural University of Medicine VOL.114;NO.1;PAGE.21-29(2005) 

Cancer prevention by astaxanthin, a natural carotenoid 

MOU X Y (Kyoto Prefectural Univ. Medicine Graduate School Of Medical Sci.) 

Astaxanthin is a natural carotenoid. The anticarcinogenic effect of astaxanthin 

was shown in mouse lung and liver models. The effect of astaxanthin on cell 

proliferation, cell cycle progression and apoptosis was examined in the HepG2 

human liver cancer cell line. Astaxanthin significantly inhibited the proliferation 

of liver cancer cells in a dose-dependent manner. Flow cytometric analysis 

demonstrated that astaxanthin restrained the cell cycle progression at G1, and 

induced apoptosis. Further examinations by real-time quantitative RT-PCR 

revealed that astaxanthin enhanced the expression of p21CIP1/WAF1, GADD153 

and c-myc genes. These results suggest that astaxanthin will be a promising agent 

for use in chemopreventive or therapeutics against cancer. 


239

Lee, S et al. (1998). J Kor Soc food Sci Nutr 27(1): 163-167, 1998. 

Language: Korean 

Inhibition of Sarcoma-180 Cell-induced Mouse Ascites Cancer by 

Astaxanthin-containing Egg Yolk 

Sang-Ho Lee, Cherl-Woo Park, Kyung-Ah Park, Young-Choon Lee, Eui-Sung 

Choi, Yeong Lae Ha 


Anticarcinogenic activity of astaxanthin-containing egg yolk(designate AEY) was 

investigated for mouse ascites carcinogenesis induced by mouse Sarcomll-I8()(S- 

I80) cells. Female ICR mice (8 mice/treatment, 7~8 weeks of age, 25±1g) were 

injected, i.p. with S-180 cells (l x 10^7 cell/ml PBS). Two days later, each mouse 

was given 0.lml PBS containing AEY(10, 25 or 50µg/g body weight) or control 

egg yolk (CEY: 50µg/g body weight) every other day for 7 times. Control mice 

were only given 0.lml S-180 cells and 0.lml PBS. Mice treated with 25µg/g body 

weight of AEY showed 24.8 days of life, which was equivalent to 138% of 

control mice's life (l8.0 days). Based on dose-dependant experiment of AEY, 

mice treated with 10µg/g body weight showed slightly longer life (l9.4 days) 

relative to mice treated with control mice, and mice treated with 50µg/g body 

weight exhibited 21.9 days of life. Mice treated with any dose of AEY exhibited 

longer life than mice with CEY 50µg/g body weight. Body weight of mice treated 

with AEY was reduced relative to that of control mice or CEY-treated mice. 

These results suggest that AEY inhibits the carcinogenesis of mouse ascites 

induced by S-180 cells. 


240

Pure & Appl. Chem. 71, 2273 (1999). 

Cancer prevention by carotenoids 

Nishino, et al, 

A review with 13 refs. Various natural carotenoids have been proven to have 

anticarcinogenic activity. Epidemiol. investigations have shown that cancer risk 

is inversely related to the consumption of green and yellow vegetables and fruits. 

As b-carotene is present in abundance in these vegetables and fruits, it has been 

investigated extensively as a possible cancer preventive agent. However, various 

carotenoids which coexist with b-carotene in vegetables and fruits also have 

anticarcinogenic activity, and some of these, such as a-carotene, lutein and 

lycopene, show a higher potency than b-carotene in suppressing exptl. 

carcinogenesis. Thus, we have carried out more extensive studies on cancer 

preventive activities of natural carotenoids in foods. For example, we found that 

b-cryptoxanthin showed antitumor initiating activity, as well as antitumor 

promoting activity. It is of interest that not only carotenoids distributed in 

vegetables and fruits, but also animal carot enoids, such as astaxanthin, are 

promising as cancer preventive agents. In the present study, the cancer preventive 

potential of phytoene was also confirmed. The establishment of NIH3T3 cells 

that produce phytoene by introducing the crtB gene provides evidence that 

resistance against transformation, imposed by transfection of activated H-ras 

oncogene, was acquired by phytoene prodn. Anal. of the action mechanism of 

these natural carotenoids is now in progress, and some interesting results have 

already been obtained; for example, various carotenoids were suggested to 

stimulate the expression of RB gene, an antioncogene. 


241

Phytopharmaceuticals in Cancer Chemoprevention CRC press D Bagchi and H. 

Preuss Ed. 2005. 

Astaxanthin and Cancer Chemoprevention 

John E. Dore, Ph.D. 

Cyanotech Corporation, Kailua-Kona, Hawaii, USA 

Introduction 

There are clear links between human cancers and diet.1,2 By some estimates, 

dietary risk 

factors rank higher than tobacco usage and much higher than pollution or 

occupational hazards in their association with cancer deaths.3 In addition to 

avoidance of tobacco smoke and carcinogenic food items, regular intake of 

chemopreventive compounds is a promising approach for reducing cancer 

incidence.3,4 A number of substances naturally occurring in foodstuffs, 

particularly antioxidant compounds in plant products, have shown promise as 

potential chemopreventive agents.3-6 Among these phytonutrients, the yellow, 

orange and red carotenoid pigments have recently sparked much interest. In 

epidemiological studies, vegetable and fruit consumption has consistently been 

associated with reduced incidence of various cancers, 5-7 and dietary carotenoid 

intake from these sources has similarly been correlated with reduced cancer 

risk.8-10 However, several recent large-scale intervention trials failed to find any 

chemopreventive effect of long-term supplementation with β-carotene, the most 

abundant dietary carotenoid.11-13 Several naturally occurring carotenoids other 

than β-carotene have exhibited anticancer activity,14-17 and are being considered 

further as potential chemopreventive agents. Among these carotenoids, the red 

pigment astaxanthin is of particular interest in health 

management due to its unique structural and chemical properties.18-20 This 

chapter will review the evidence for anticarcinogenic behavior of selected 

carotenoids, with an emphasis on the chemopreventive activities of astaxanthin. 


242

J Herb Pharmacother. 2005;5(1):17-26. 

A preliminary investigation of the enzymatic inhibition of 5alphareduction 

and growth of prostatic carcinoma cell line LNCap-FGC 

by natural astaxanthin and Saw Palmetto lipid extract in vitro. 

Anderson ML. 

Inhibition of 5alpha-reductase has been reported to decrease the symptoms of 

benign prostate hyperplasia (BPH) and possibly inhibit or help treat prostate 

cancer. Saw Palmetto berry lipid extract (SPLE) is reported to inhibit 5alphareductase 

and decrease the clinical symptoms of BPH. Epidemiologic studies 

report that carotenoids such as lycopene may inhibit prostate cancer. In this 

investigation the effect of the carotenoid astaxanthin, and SPLE were examined 

for their effect on 5alpha-reductase inhibition as well as the growth of prostatic 

carcinoma cells in vitro. The results show astaxanthin demonstrated 98% 

inhibition of 5alpha-reductase at 300 microg/mL in vitro. Alphastat, the 

combination of astaxanthin and SPLE, showed a 20% greater inhibition of 5alphareductase 

than SPLE alone n vitro. CONCLUSIONS: Low levels of carotenoid 

astaxanthin inhibit 5alpha-reductase and decrease the growth of human prostatic 

cancer cells in vitro. Astaxanthin added to SPLE shows greater inhibition of 

5alpha-reductase than SPLE alone in vitro. 


243

Invest New Drugs. 2009 Oct 30. [Epub ahead of print] 

Astaxanthin inhibits tumor invasion by decreasing extracellular matrix 

production and induces apoptosis in experimental rat colon 

carcinogenesis by modulating the expressions of ERK-2, NFkB and 

COX-2. 

Nagendraprabhu P, Sudhandiran G. 

Department of Biochemistry, University of Madras, Guindy campus, Chennai, 600 025, 

Tamil nadu, India. 


Colon cancer is the third most malignant neoplasm in the world and it remains an 

important cause of mortality in Asian and Western countries. Astaxanthin (AST), a major 

component of carotenoids possesses attractive remedial features. The purpose of this 

study is to investigate the possible mechanism of action of astaxanthin against 1, 2 

dimethyl hydrazine (DMH)-induced rat colon carcinogenesis. Wistar male rats were 

randomized into five groups, group 1 were control rats, group 2 were rats that received 

AST (15 mg/kg body wt p.o. everyday), rats in group 3 were induced with DMH (40 

mg/kg body wt, s.c.), DMH-induced rats in groups 4 and 5 were either pre or post 

initiated with AST, respectively as in group 2. DMH-induced rats exhibited elevated 

expressions of Nuclear factor kappa B-p65 (NF-kappaB-p65), Cyclooxygenase-2 (COX- 

2), Matrixmetallo proteinases (MMP) 2/9, Proliferating cell nuclear antigen (PCNA), and 

Extracellular signal-regulated kinase-2 (ERK-2) as confirmed by immunofluorescence. 

Further, Westernblot analysis of MMPs-2/9, ERK-2 and Protein kinase B (Akt) revealed 

increased expressions of these proteins in DMH-induced groups of rats. AST-treatment 

decreased the expressions of all these vital proteins, involved in colon carcinogenesis. 

The ability of AST to induce apoptosis in the colon of DMH-induced rats was confirmed 

by Annexin-V/PI staining in a confocal microscopy, DNA fragmentation analysis and 

expression of caspase-3 by Western blotting. In conclusion, astaxanthin exhibits antiinflammatory 

and anti-cancer effects by inducing apoptosis in DMH-induced rat colon 

244

carcinogenesis by modulating the expressions of NFkB, COX-2, MMPs-2/9, Akt and 

ERK-2. 



245

Chem Biol Interact. 2009 Aug 14;180(3):398-406. Epub 2009 Apr 2. 

Intervention of astaxanthin against cyclophosphamide-induced 

oxidative stress and DNA damage: a study in mice. 



Education and Research, Sector-67, S.A.S. Nagar, Punjab 160062, India. 


Astaxanthin, a natural and nutritional red carotenoid pigment, is used as a dietary 

supplement. The intention of the present study was to investigate the beneficial effects of 

dietary pigment astaxanthin, against cyclophosphamide-induced oxidative stress and 

DNA damage. The end points of evaluation of the study included: (a) malondialdehyde, 

glutathione and superoxide dismutase concentration in liver to detect oxidative stress; (b) 

normal and modified alkaline comet assays (the latter includes lesion-specific enzymes 

formamidopyrimidine-DNA glycosylase and endonuclease-III) to detect normal and 

oxidative stress-induced DNA damage by cyclophosphamide in the mouse bone marrow 

and the peripheral blood lymphocytes. In addition, micronucleus assay and chromosomal 

aberration test capable of detecting the DNA damage were also carried out in peripheral 

blood and bone marrow of mice. Cyclophosphamide (100 mg/kg intra-peritoneal) 

treatment led to significant increase in liver malondialdehyde and decreased the 

antioxidant enzymes glutathione and superoxide dismutase. Further, cyclophosphamide 

also significantly increased the DNA damage as observed from normal and modified 

comet assays as well as micronucleus and chromosomal aberration assay. Pre-treatment 

with astaxanthin (12.5, 25 and 50 mg/kg/day for 5 days per oral) resulted in the 

restoration of oxidative stress markers such as malondialdehyde, glutathione and 

superoxide dismutase in liver. The amelioration of oxidative stress with astaxanthin pretreatment 

correlated well with the decreased DNA damage as evident from normal and 

modified alkaline comet assays of bone marrow cells and peripheral blood lymphocytes. 

Further astaxanthin pre-treatment also reduced the frequency of chromosomal breakage 

and micronucleus formation in the mouse bone marrow cells and peripheral blood 

reticulocytes. It is thus concluded that pre-treatment with astaxanthin attenuates 

cyclophosphamide-induced oxidative stress and subsequent DNA damage in mice and it 

can be used as a chemoprotective agent against the toxicity of anticancer drug 

cyclophosphamide. 



246

Cancer Lett. 2009 Sep 28;283(1):108-17. Epub 2009 May 6. 

Growth-inhibitory effects of the astaxanthin-rich alga Haematococcus 

pluvialis in human colon cancer cells. 

Palozza P, Torelli C, Boninsegna A, Simone R, Catalano A, Mele MC, Picci N. 

Institute of General Pathology, Catholic University School of Medicine, L. Go F. Vito, 1 

00168 Rome, Italy. p.palozza@rm.unicatt.it 


The growth-inhibitory effects of the astaxanthin-rich Haematococcus pluvialis were 

studied in HCT-116 colon cancer cells. H. pluvialis extract (5-25 microg/ml) inhibited 

cell growth in a dose- and time-dependent manner, by arresting cell cycle progression and 

by promoting apoptosis. At 25 microg/ml of H. pluvialis extract, an increase of p53, 

p21(WAF-1/CIP-1) and p27 expression (220%, 160%, 250%, respectively) was 

observed, concomitantly with a decrease of cyclin D1 expression (58%) and AKT 

phosphorylation (21%). Moreover, the extract, at the same concentration, strongly upregulated 

apoptosis by modifying the ratio of Bax/Bcl-2 and Bcl-XL, and increased the 

phosphorylation of p38, JNK, and ERK1/2 by 160%, 242%, 280%, respectively. Growthinhibitory 

effects by H. pluvialis were also observed in HT-29, LS-174, WiDr, SW-480 

cells. This study suggests that H. pluvialis may protect from colon cancer. 



247

Fundam Clin Pharmacol. 2009 Apr;23(2):225-34. 

Antioxidative and antiproliferative effects of astaxanthin during the 

initiation stages of 1,2-dimethyl hydrazine-induced experimental colon 

carcinogenesis. 

Prabhu PN, Ashokkumar P, Sudhandiran G. 

Department of Biochemistry, University of Madras, Guindy campus, Chennai - 600 025, 

Tamil Nadu, India. 


Colon cancer is one of the major causes of cancer mortality worldwide. Several 

carotenoids with antioxidant properties are reported for their chemopreventive nature. In 

this study, we have evaluated the chemopreventive efficacy of astaxanthin on lipid 

peroxidation, antioxidant status, total number of aberrant crypt foci (ACF), and cell 

proliferation in 1,2 dimethylhydrazine (DMH)-induced colon carcinogenesis using a rat 

model. DMH was induced subcutaneously at a dosage of 40 mg/kg body weight, twice a 

week for 2 weeks. Astaxanthin was administered before and after the DMH induction, 

orally at a concentration of 15 mg/kg body weight throughout the experimental period. At 

the end of 16 weeks, pre-treatment with astaxanthin markedly reduced the degree of 

histological lesions, ACF development and also lowered the number of argyrophilic 

nucleolar organizer regions. Our results also showed the decreased levels of colon 

enzymic and non-enzymic antioxidants and increased levels of lipid peroxidation marker 

levels in DMH-induced rats, which were significantly reversed on astaxanthin 

administration. In conclusion, the results of this study suggest that astaxanthin has an 

affirmative and beneficial effect against chemically induced colonic pre-neoplastic 

progression in rats induced by DMH. 



248

Anticancer Res. 2010 Jun;30(6):2171-5. 

Effect of dietary astaxanthin at different stages of mammary tumor 

initiation in BALB/c mice. 

Nakao R, Nelson OL, Park JS, Mathison BD, Thompson PA, Chew BP. 

School of Food Science, Washington State University, Pullman, WA 99164, USA. 


The effects of astaxanthin on tumor growth, cardiac function and immune response in 

mice were studied. Female BALB/c mice were fed a control diet (diet C) for 8 weeks, 

0.005% astaxathin for 8 weeks (diet A), or diet C for weeks 1-5 followed by diet A 

thereafter (diet CA). Mice were injected with a mammary tumor cell line on day 7 and 

tumor growth was measured daily. Mice fed diet A had extended tumor latency and lower 

tumor volume (p

Invest New Drugs. 2011 Apr;29(2):207-24. Epub 2009 Oct 30. 

Astaxanthin inhibits tumor invasion by decreasing 

extracellular matrix production and induces apoptosis in 

experimental rat colon carcinogenesis by modulating the 

expressions of ERK-2, NFkB and COX-2. 

Nagendraprabhu P, Sudhandiran G. 

Source 

Department of Biochemistry, University of Madras, Chennai, Tamil nadu, India. 


Colon cancer is the third most malignant neoplasm in the world and it remains an 

important cause of mortality in Asian and Western countries. Astaxanthin (AST), a major 

component of carotenoids possesses attractive remedial features. The purpose of this 

study is to investigate the possible mechanism of action of astaxanthin against 1, 2 

dimethyl hydrazine (DMH)-induced rat colon carcinogenesis. Wistar male rats were 

randomized into five groups, group 1 were control rats, group 2 were rats that received 

AST (15 mg/kg body wt p.o. everyday), rats in group 3 were induced with DMH (40 

mg/kg body wt, s.c.), DMH-induced rats in groups 4 and 5 were either pre or post 

initiated with AST, respectively as in group 2. DMH-induced rats exhibited elevated 

expressions of Nuclear factor kappa B-p65 (NF-κB-p65), Cyclooxygenase-2 (COX-2), 

Matrixmetallo proteinases (MMP) 2/9, Proliferating cell nuclear antigen (PCNA), and 

Extracellular signal-regulated kinase-2 (ERK-2) as confirmed by immunofluorescence. 

Further, Westernblot analysis of MMPs-2/9, ERK-2 and Protein kinase B (Akt) revealed 

increased expressions of these proteins in DMH-induced groups of rats. AST-treatment 

decreased the expressions of all these vital proteins, involved in colon carcinogenesis. 

The ability of AST to induce apoptosis in the colon of DMH-induced rats was confirmed 

by Annexin-V/PI staining in a confocal microscopy, DNA fragmentation analysis and 

expression of caspase-3 by Western blotting. In conclusion, astaxanthin exhibits antiinflammatory 

and anti-cancer effects by inducing apoptosis in DMH-induced rat colon 

carcinogenesis by modulating the expressions of NFkB, COX-2, MMPs-2/9, Akt and 

ERK-2. 



250

Biol Pharm Bull. 2011;34(6):839-44. 

Astaxanthin induces mitochondriamediated 

apoptosis in rat hepatocellular 

carcinoma CBRH-7919 cells. 

Song XD, Zhang JJ, Wang MR, Liu WB, Gu XB, Lv CJ. 

Source 

Medicine Research Center, Binzhou Medical University, Yantai, China. 


We designed to study the role of mitochondria in astaxanthin-induced apoptosis in 

hepatocellular carcinoma cells. Effect of astaxanthin on cell proliferation was studied by 

using methyl thiazolyl tetrazolium (MTT) in three tumor cell lines (CBRH-7919, SHZ-88 

and Lewis) and normal human hepatocyte HL-7702 cell. Cell apoptosis rate, changes of 

mitochondrial morphology, mitochondrial transmembrane potential and electron transport 

chain were evaluated respectively. Expressions of B cell lymphoma/leukemia-2 (Bcl-2) 

and Bcl-2 associated X protein (Bax) were detected by Western blot. Results as 

following, astaxanthin had little effect on HL-7702 cell, however its inhibition was most 

pronounced in CBRH-7919 cell line with an IC₅₀ of 39 µM. This dose of astaxanthin and 

CBRH-7919 cell line were chosen for further studies. Astaxanthin could induce cell 

apoptosis and mitochondrial membrane damage. The mitochondrial transmembrane 

potential and function of electron transport chain were decreased. The expression of Bcl- 

2 protein was down-regulated but that of Bax protein was up-regulated. In conclusion, 

astaxanthin showed anticancer effect by inducing cell apoptosis through the regulation of 

mitochondrial-dependent manner. 



251

Am J Vet Res. 2010 Jan;71(1):89-96. 

Evaluation of the protective effects of all-trans-astaxanthin on 

canine osteosarcoma cell lines. 

Wakshlag JJ, Balkman CA, Morgan SK, McEntee MC. 

Source 

Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, 

Ithaca, NY 14853, USA. jw37@cornell.edu 


OBJECTIVE: To determine the effects of the antioxidant astaxanthin on growth of 

canine osteosarcoma cells with and without concurrent chemotherapeutic or irradiation 

insult. 

SAMPLE POPULATION: Cells from 3 established canine osteosarcoma cell lines (D17, 

OS 2.4, and HMPOS). 

PROCEDURES: Growth-curve kinetics and cell cytotoxic effects were assessed by 

means of various treatment combinations and a 3-(4,5-dimethylthiazol-2-yl)-2,5- 

diphenyltetrazolium bromide assay. Western blotting was performed to examine 

previously identified signaling pathways that astaxanthin reportedly affects. Additionally, 

cell-cycle kinetic evaluations, soft agar colony-forming assays, and antioxidant assays 

were performed to better understand the effect of astaxanthin on cell growth and function. 

RESULTS: Exposure to astaxanthin alone resulted in a mild to pronounced attenuation of 

cell proliferation in vitro, depending on the cell line, and did not interfere with the celldeath 

response to doxorubicin, irradiation, or peroxide-mediated insult. In some 

instances, astaxanthin acted in an additive fashion to augment cell death. Astaxanthin 

exposure increased the antioxidant potential of cells, whereas peroxide-mediated cell 

stress increased the antioxidant potential to the same degree as astaxanthin exposure or 

greater. No dramatic changes in phosphorylation of protein kinase B or upregulation of 

connexin 43 were detected. 

CONCLUSIONS AND CLINICAL RELEVANCE: Findings suggested that astaxanthin 

administration may be beneficial in treatment of dogs for osteosarcoma. Its actions as an 

antioxidant did not improve osteosarcoma cell survival during chemotherapeutic or 

irradiation insults, warranting further research into this natural compound as an adjuvant, 

antiproliferative treatment for osteosarcoma in dogs. 



252

Diabetes 

Life Sci. 2007 Jan 16;80(6):522-9. Epub 2006 Oct 12. 

Astaxanthin ameliorates features of metabolic syndrome in 

SHR/NDmcr-cp. 

Hussein G, Nakagawa T, Goto H, Shimada Y, Matsumoto K, Sankawa U, 

Watanabe H. 

International Research Center for Traditional Medicine, Toyama, Toyama 

Prefecture 939-8224, Japan. ghazihussein@hotmail.com 

Glucose and lipid metabolic parameters play crucial roles in metabolic syndrome 

and its major feature of insulin resistance. This study was designed to investigate 

whether dietary astaxanthin oil (ASX-O) has potential effects on metabolic 

syndrome features in an SHR/NDmcr-cp (cp/cp) rat model. Oral administration of 

ASX (50 mg/kg/day) for 22 weeks induced a significant reduction in arterial 

blood pressure in SHRcp. It also significantly reduced the fasting blood glucose 

level, homeostasis index of insulin resistance (HOMA-IR), and improved insulin 

sensitivity. The results also showed an improved adiponectin level, a significant 

increase in high-density lipoprotein cholesterol, a significant decrease in plasma 

levels of triglycerides, and non-esterified fatty acids. Additionally, ASX showed 

significant effects on the white adipose tissue by decreasing the size of the fat 

cells. These results suggest that ASX ameliorates insulin resistance by 

mechanisms involving the increase of glucose uptake, and by modulating the level 

of circulating lipid metabolites and adiponectin. 



Diabetes 

253

J Cell Biochem. 2008 Apr 15;103(6):1925-37. 

Astaxanthin protects mesangial cells from hyperglycemia-induced 

oxidative signaling. 

Manabe E, Handa O, Naito Y, Mizushima K, Akagiri S, Adachi S, Takagi T, 

Kokura S, Maoka T, Yoshikawa T. 

School of Nursing, Kyoto Prefectural University of Medicine, Kyoto 602-8566, 

Japan. 

Astaxanthin (ASX) is a carotenoid that has potent protective effects on diabetic 

nephropathy in mice model of type 2 diabetes. In this study, we investigated the 

protective mechanism of ASX on the progression of diabetic nephropathy using 

an in vitro model of hyperglycemia, focusing on mesangial cells. Normal human 

mesangial cells (NHMCs) were cultured in the medium containing normal (5 

mM) or high (25 mM) concentrations of D-glucose. Reactive oxygen species 

(ROS) production, the activation of nuclear transcription factors such as nuclear 

factor kappa B (NFkappaB) and activator protein-1 (AP-1), and the 

expression/production of transforming growth factor-beta 1 (TGFbeta(1)) and 

monocyte chemoattractant protein-1 (MCP-1) were evaluated in the presence or 

absence of ASX. High glucose (HG) exposure induced significant ROS 

production in mitochondria of NHMCs, which resulted in the activation of 

transcription factors, and subsequent expression/production of cytokines that 

plays an important role in the mesangial expansion, an important event in the 

pathogenesis of diabetic nephropathy. ASX significantly suppressed HG-induced 

ROS production, the activation of transcription factors, and cytokine 

expression/production by NHMCs. In addition, ASX accumulated in the 

mitochondria of NHMCs and reduced the production of ROS-modified proteins in 

mitochondria. ASX may prevent the progression of diabetic nephropathy mainly 

through ROS scavenging effect in mitochondria of mesangial cells and thus is 

expected to be very useful for the prevention of diabetic nephropathy. 


Diabetes 

254

Int J Vitam Nutr Res. 2008 Jul-Sep;78(4-5):175-82. 

Inhibitory effect of astraxanthin combined with Flavangenol on 

oxidative stress biomarkers in streptozotocin-induced diabetic rats. 

Nakano M, Orimo N, Katagiri N, Tsubata M, Takahashi J, Van Chuyen N. 

Department of Food and Nutrition, Japan Women's University, Tokyo, Japan. 

masako.nakano06@gr.jwu.ac.jp 

In this study, the effect of dietary antioxidants, such as astaxanthin and 

Flavangenol, and a combination of both, in counteracting oxidative stress in 

streptozotocin-induced diabetes was investigated. Streptozotocin-induced diabetic 

rats were divided into four groups: control, astaxanthin, Flavangenol, and 

combined astaxanthin and Flavangenol (mix group). Each group other than the 

control group was fed with an astaxanthin diet (0.1 g/kg), Flavangenol diet (2.0 

g/kg), or an astaxanthin (0.1 g/kg)-Flavangenol (2.0 g/kg) mixture diet, 

respectively. After 12 weeks of feeding, the results showed that the lipid peroxide 

levels of plasma and lens and the plasma triglyceride (TG) level in the mix group 

were significantly decreased by 44%, 20%, and 20%, respectively, compared with 

the control group. In the mix group, lipid peroxidation was also significantly 

reduced by 70% in the liver and 20% in the kidney compared with the control 

group. Furthermore, the level of urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) 

in the mix group was significantly lower, 36%, than the control group. The alphatocopherol 

concentrations in the plasma, liver, and kidney in the astaxanthin and 

mix groups were significantly higher, 3-9 times, than in the control group. The 

degree of cataract formation in the Flavangenol and mix groups tended to be 

lower than the control group. These results indicate that the combination of 

astaxanthin with Flvangenol has an improved protective effect on oxidative stress 

associated with streptozotocin-induced diabetes than either agent used alone. 

Thus, this combination may be beneficial in preventing the progression of diabetic 

complications. 


Diabetes 

255

J Nutr Sci Vitaminol (Tokyo). 2008 Aug;54(4):329-34. 

Effect of astaxanthin in combination with alpha-tocopherol or 

ascorbic acid against oxidative damage in diabetic ODS rats. 

Nakano M, Onodera A, Saito E, Tanabe M, Yajima K, Takahashi J, Nguyen 

VC. 

Department of Food and Nutrition, Japan Women's University Japan Women's 

University Tokyo 112-8681, Japan. masako.nakano06@gr.jwu.ac.jp 

The present study was performed to investigate the effect of astaxanthin in 

combination with other antioxidants against oxidative damage in streptozotocin 

(STZ)-induced diabetic Osteogenic Disorder Shionogi (ODS) rats. Diabetic-ODS 

rats were divided into five groups: control, astaxanthin, ascorbic acid, alphatocopherol, 

and tocotrienol. Each of the four experimental groups was 

administered a diet containing astaxanthin (0.1 g/kg), in combination with 

ascorbic acid (3.0 g/kg), alpha-tocopherol (0.1 g/kg), or tocotrienol (0.1 g/kg) for 

20 wk. The effects of astaxanthin with other antioxidants on lipid peroxidation, 

urinary 8-hydroxy-2-deoxyguanosine (8-OHdG) excretion, serum creatinine (Cr) 

level, creatinine clearance (Ccr), and urinary protein content were assessed. The 

serum lipid peroxide levels and chemiluminescent (CL) intensity in the liver of 

the alpha-tocopherol and tocotrienol groups were significantly reduced in 

comparison to that of the control group. In the alpha-tocopherol group, urinary 8- 

OHdG excretion, serum Cr level, Ccr, urinary albumin excretion, and urinary 

protein concentration were significantly decreased as compared with those in the 

control group. Additionally, the CL intensity in the kidney of the alpha-tocopherol 

group was significantly lower, but that of the ascorbic acid group was 

significantly higher than that in the control group. These results indicate that 

dietary astaxanthin in combination with alpha-tocopherol has an inhibitory effect 

on oxidative stress. On the other hand, our study suggests that excessive ascorbic 

acid intake increases lipid peroxidation in diabetic rats. 


Diabetes 

256

Int J Mol Med. 2006 Oct;18(4):685-95. 

Microarray profiling of gene expression patterns in glomerular cells of 

astaxanthin-treated diabetic mice: a nutrigenomic approach. 

Naito Y, Uchiyama K, Mizushima K, Kuroda M, Akagiri S, Takagi T, Handa O, 

Kokura S, Yoshida N, Ichikawa H, Takahashi J, Yoshikawa T. 

Department of Medical Proteomics, Kyoto Prefectural University of Medicine, Kyoto 

602-8566, Japan. ynaito@koto.kpu-m.ac.jp 

We have demonstrated that astaxanthin reduces glomerular oxidative stress as well as 

inhibits the increase in urinary albumin in diabetic db/db mice. The aim of the present 

study was to determine the gene expression patterns in the glomerular cells of the diabetic 

mouse kidney, and to investigate the effects of astaxanthin on the expression of these 

genes using a high-density DNA microarray. The diet administered to the astaxanthinsupplementation 

group was prepared by mixing a control powder with astaxanthin at a 

concentration of 0.02%. Glomerular cells were obtained from the kidneys of mice by 

laser capture microdissection. Preparation of cRNA and target hybridization were 

performed according to the Affymetrix GeneChip eukaryotic small sample target labeling 

assay protocol. The gene expression profile was evaluated by the mouse expression set 

430A GeneChip. Array data analysis was carried out using Affymetrix GeneChip 

operating and Ingenuity Pathway analysis software. Comparison between diabetic db/db 

and non-diabetic db/m mice revealed that 779 probes (3.1%) were significantly affected, 

i.e. 550 probes were up-regulated, and 229 probes were down-regulated, both at levels of 

>/=1.5-fold in the diabetic mice. Ingenuity signal analysis of 550 up-regulated probes 

revealed the mitochondrial oxidative phosphorylation pathway as the most significantly 

affected caronical pathway. The affected genes were associated with complexes I, III, and 

IV located on the mitochondrial inner membrane, and the expression levels of these genes 

were decreased in mice treated with astaxanthin as compared to the levels in the control 

mice. In addition, the expression of many genes associated with oxidative stress, collagen 

synthesis, and transforming growth factor-beta signaling was enhanced in the diabetic 

mice, and this enhancement was slightly inhibited in the astaxanthin-treated mice. In 

conclusion, this genome-wide nutrigenomics approach provided insight into genes and 

putative genetic pathways that are thought to be affected by stimulation by high-glucose 

concentrations. In addition, the present approach may help us gain a better understanding 

of the genes and pathways involved in the anti-diabetic mechanism of astaxanthin. 



Diabetes 

257

Biofactors. 2004;20(1):49-59. 

Prevention of diabetic nephropathy by treatment with astaxanthin 

in diabetic db/db mice. 

Naito Y, Uchiyama K, Aoi W, Hasegawa G, Nakamura N, Yoshida N, Maoka 

T, Takahashi J, Yoshikawa T. 

Molecular Gastroenterology and Hepatology, Graduate School of Medical 

Science, Kyoto Prefectural University of Medicine, Kamigyo-ku, Kyoto 602- 

8566, Japan. 

Oxidative stress is implicated as an important mechanism by which diabetes 

causes nephropathy. Astaxanthin, which is found as a common pigment in algae, 

fish, and birds, is a carotenoid with significant potential for antioxidative activity. 

In this study, we examined whether chronic administration of astaxanthin could 

prevent the progression of diabetic nephropathy induced by oxidative stress in 

mice. We used female db/db mice, a rodent model of type 2 diabetes, and their 

non-diabetic db/m littermates. The mice were divided into three groups as 

follows: non-diabetic db/m, diabetic db/db, and diabetic db/db treated with 

astaxanthin. Blood glucose level, body weight, urinary albumin, and urinary 8- 

hydroxydeoxyguanosine (8-OHdG) were measured during the experiments. 

Histological and 8-OHdG immunohistochemical studies were performed for 12 

weeks from the beginning of treatment. After 12 weeks of treatment, the 

astaxanthin-treated group showed a lower level of blood glucose compared with 

the non-treated db/db group; however, both groups had a significantly high level 

compared with the db/m mice. The relative mesangial area calculated by the 

mesangial area/total glomerular area ratio was significantly ameliorated in the 

astaxanthin-treated group compared with the non-treated db/db group. The 

increases in urinary albumin and 8-OHdG at 12 weeks of treatment were 

significantly inhibited by chronic treatment with astaxanthin. The 8-OHdG 

immunoreactive cells in glomeruli of non-treated db/db mice were more 

numerous than in the astaxanthin-treated db/db mice. In this study, treatment with 

astaxanthin ameliorated the progression and acceleration of diabetic nephropathy 

in the rodent model of type 2 diabetes. The results suggested that the antioxidative 

activity of astaxanthin reduced the oxidative stress on the kidneys and prevented 

renal cell damage. In conclusion, administration of astaxanthin might be a novel 

approach for the prevention of diabetes nephropathy. 


Diabetes 

258

Redox Rep. 2002;7(5):290-3. 

Astaxanthin protects beta-cells against glucose toxicity in diabetic 

db/db mice. 

Uchiyama K, Naito Y, Hasegawa G, Nakamura N, Takahashi J, Yoshikawa 

T. 

First Department of Medicine, Kyoto Prefectural University of Medicine, Kyoto, 

Japan. 

Oxidative stress induced by hyperglycemia possibly causes the dysfunction of 

pancreatic beta-cells and various forms of tissue damage in patients with diabetes 

mellitus. Astaxanthin, a carotenoid of marine microalgae, is reported as a strong 

anti-oxidant inhibiting lipid peroxidation and scavenging reactive oxygen species. 

The aim of the present study was to examine whether astaxanthin can elicit 

beneficial effects on the progressive destruction of pancreatic beta-cells in db/db 

mice--a well-known obese model of type 2 diabetes. We used diabetic 

C57BL/KsJ-db/db mice and db/m for the control. Astaxanthin treatment was 

started at 6 weeks of age and its effects were evaluated at 10, 14, and 18 weeks of 

age by non-fasting blood glucose levels, intraperitoneal glucose tolerance test 

including insulin secretion, and beta-cell histology. The non-fasting blood glucose 

level in db/db mice was significantly higher than that of db/m mice, and the 

higher level of blood glucose in db/db mice was significantly decreased after 

treatment with astaxanthin. The ability of islet cells to secrete insulin, as 

determined by the intraperitoneal glucose tolerance test, was preserved in the 

astaxanthin-treated group. Histology of the pancreas revealed no significant 

differences in the beta-cell mass between astaxanthin-treated and -untreated db/db 

mice. In conclusion, these results indicate that astaxanthin can exert beneficial 

effects in diabetes, with preservation of beta-cell function. This finding suggests 

that anti-oxidants may be potentially useful for reducing glucose toxicity. 


Diabetes 

259

Redox Report, Vol. 7, No. 5, 2002 290-3 

Astaxanthin protects β-cells against glucose toxicity in diabetic 

db/db mice 

Kazuhiko Uchiyama1, Yuji Naito1, Goji Hasegawa1, Naoto Nakamura1, 

Jiro Takahashi2, Toshikazu Yoshikawa1 

Oxidative stress induced by hyperglycemia possibly causes the dysfunction of 

pancreatic β–cells and various forms of tissue damage in patients with diabetes 

mellitus. Astaxanthin, a carotenoid of marine microalgae, is reported as a strong 

anti-oxidant inhibiting lipid peroxidation and scavenging reactive oxygen species. 

The aim of the present study was to examine whether astaxanthin can elicit 

beneficial effects on the progressive destruction of pancreatic -cells in db/db mice 

β – a well-known obese model of type 2 diabetes. We used diabetic C57BL/KsJdb/db 

mice and db/m for the control. Astaxanthin treatment was started at 6 

weeks of age and its effects were evaluated at 10, 14, and 18 weeks of age by 

non-fasting blood glucose levels, intraperitoneal glucose tolerance test including 

insulin secretion, and -cell histology. The non-fasting blood glucose level in 

db/db mice was significantly higher than that of db/m mice, and the higher level 

of blood glucose in db/db mice was significantly decreased after treatment with 

astaxanthin. The ability of islet cells to secrete insulin, as determined by the 

intraperitoneal glucose tolerance test, was preserved in the astaxanthin-treated 

group. Histology of the pancreas revealed no significant differences in the -cell 

mass between astaxanthin-treated and -untreated db/db mice. In conclusion, these 

results indicate that astaxanthin can exert beneficial effects in diabetes, with 

preservation of β-cell function. This finding suggests that anti-oxidants may be 

potentially useful for reducing glucose toxicity. 

Diabetes 

260

Chem Biol Interact. 2010 Aug 5;186(3):306-15. Epub 2010 May 31. 

Astaxanthin ameliorates the redox imbalance in lymphocytes of 

experimental diabetic rats. 

Otton R, Marin DP, Bolin AP, Santos Rde C, Polotow TG, Sampaio SC, de Barros MP. 

Postgraduate Program, Human Movement Sciences, Institute of Physical Activity and 

Sport Sciences, Universidade Cruzeiro do Sul, ZIP 01506000, Sao Paulo, SP, Brazil; 

Postgraduate Program, Health Sciences, CBS, Universidade Cruzeiro do Sul, ZIP 

08060070, Sao Paulo, SP, Brazil. 


Diabetes mellitus is a syndrome of impaired insulin secretion/sensitivity and frequently 

diagnosed by hyperglycemia, lipid abnormalities, and vascular complications. The 

diabetic 'glucolipotoxicity' also induces immunodepression in patients by redox 

impairment of immune cells. Astaxanthin (ASTA) is a pinkish-orange carotenoid found 

in many marine foods (e.g. shrimp, crabs, salmon), which has powerful antioxidant, 

photoprotective, antitumor, and cardioprotective properties. Aiming for an antioxidant 

therapy against diabetic immunodepression, we here tested the ability of prophylactic 

ASTA supplementation (30 days, 20 mg ASTA/kg BW) to oppose the redox impairment 

observed in isolated lymphocytes from alloxan-induced diabetic Wistar rats. The redox 

status of lymphocytes were thoroughly screened by measuring: (i) production of 

superoxide (O(2)(-)), nitric oxide (NO), and hydrogen peroxide (H(2)O(2)); (ii) cytosolic 

Ca(2+); (iii) indexes of oxidative injury; and (iv) activities of major antioxidant enzymes. 

Hypolipidemic and antioxidant effects of ASTA in plasma of ASTA-fed/diabetic rats 

were apparently reflected in the circulating lymphocytes, since lower activities of 

catalase, restored ratio between glutathione peroxidase and glutathione reductase 

activities and lower scores of lipid oxidation were concomitantly measured in those 

immune cells. Noteworthy, lower production of NO and O(2)(-) (precursors of 

peroxynitrite), and lower cytosolic Ca(2+) indicate a hypothetical antiapoptotic effect of 

ASTA in diabetic lymphocytes. However, questions are still open regarding the proper 

ASTA supplementation dose needed to balance efficient antioxidant protection and 

essential NO/H(2)O(2)-mediated proliferative capacities of diabetic lymphocytes. 


Diabetes 

261

Int Endod J. 2010 Jun 8. [Epub ahead of print] 

In vivo astaxanthin treatment partially prevents antioxidant alterations 

in dental pulp from alloxan-induced diabetic rats. 

Leite MF, de Lima A, Massuyama MM, Otton R. 

Ciências Biológicas e da Saúde, Universidade Cruzeiro do Sul - São Paulo, Brazil. 


Leite MF, de Lima A, Massuyama MM, Otton R.In vivo astaxanthin treatment partially 

prevents antioxidant alterations in dental pulp from alloxan-induced diabetic rats. 

International Endodontic Journal. Abstract Aim To evaluate the effect of astaxanthin on 

antioxidant parameters of dental pulp from diabetic rats. The hypothesis tested was that 

supplementation of diabetic rats with astaxanthin might eliminate, or at least attenuate, 

the defect in their antioxidative status. Methodology Wistar rats (n = 32) were divided 

into four groups: untreated control, treated control, untreated diabetic and treated diabetic 

rats. A prophylactic dose of astaxanthin (20 mg kg(-1) body weight) was administered 

daily by gavage for 30 days. On day 23, diabetes was induced by injection of alloxan (60 

mg kg(-1) body weight). After 7 days of diabetes induction, the rats were killed, and pulp 

tissue from incisor teeth removed. Superoxide dismutase (SOD), catalase, glutathione 

peroxidase (GPx) and reductase activities were determined. Data were compared by 

anova and the Newman-Keuls test (P < 0.05). Results Diabetes caused a reduction in 

SOD, GPx and reductase activity in dental pulp tissue. Astaxanthin had no effect on SOD 

and catalase activities; however, it stimulated GPx in control and diabetic rats. 

Conclusions Diabetes altered the antioxidant system in dental pulp tissue; astaxanthin 

partially improved the diabetic complications. 


Diabetes 

262

Arch Oral Biol. 2010 Jul;55(7):479-85. 

Astaxanthin restores the enzymatic antioxidant profile in salivary gland 

of alloxan-induced diabetic rats. 

Leite MF, Lima AM, Massuyama MM, Otton R. 

Universidade Cruzeiro do Sul, CEP, São Paulo, Brazil. 

mariana.leite@cruzeirodosul.edu.br 


OBJECTIVE: To evaluate the effect of astaxanthin on antioxidant parameters of 

salivary gland from diabetic rats. The hypothesis of the study was whether the 

supplementation of diabetic rats with astaxanthin might antagonize, or at least prevent, 

the defect in their antioxidative status. 

DESIGN: Wistar rats (n=32) were divided in 4 groups: untreated control, treated control, 

untreated diabetic and treated diabetic rats. Astaxanthin (20mg/kg body weight) was 

administered daily by gavage for 30 days. On day 23, diabetes was induced by injection 

of alloxan (60 mg/kg body weight). After 7 days of diabetes induction, the rats were 

killed and submandibular and parotid removed. Superoxide dismutase (SOD), catalase, 

glutathione peroxidase and reductase activities and the content of thiol groups were 

determined. Data were compared by ANOVA and the Tukey test (p

J Agric Food Chem. 2009 Oct 14;57(19):8793-7. 

Protection against oxidative stress, inflammation, and apoptosis of highglucose-exposed 

proximal tubular epithelial cells by astaxanthin. 

Kim YJ, Kim YA, Yokozawa T. 

Department of Dental Hygiene, Busan Women's College, Busanjin-Gu, Busan, Korea. 


Astaxanthin is a carotenoid with powerful antioxidant properties that exists naturally in 

various plants, algae, and seafood. The purpose of the present study is to examine the 

protective action of astaxanthin against high-glucose-induced oxidative stress, 

inflammation, and apoptosis in proximal tubular epithelial cells (PTECs). To assess the 

efficacy of astaxanthin, several key markers and activities were measured, including lipid 

peroxidation, total reactive species (RS), superoxide (*O(2)), nitric oxide (NO*), and 

peroxynitrite (ONOO(-)), as well as expressions of inflammatory proteins, inducible 

nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), nuclear factor-kappa B 

(NF-kappaB) nuclear translocation, and levels of Bcl2/Bax protein. Results showed that 

astaxanthin effectively suppressed lipid peroxidation, total RS, *O(2), NO*, ONOO(-), 

iNOS and COX-2 protein levels, NF-kappaB nuclear translocation, and pro-apototic Bax, 

whereas it increased anti-apoptotic Bcl2 protein levels. On the basis of these findings, it 

was concluded that in PTECs, astaxanthin has a protective efficacy against several 

deleterious effects caused by high glucose exposure and proposed that astaxanthin should 

be explored further as a potential antidiabetic remedy for the treatment of diabetic 

nephropathy. 


Diabetes 

264


Ameliorative effect of astaxanthin on 

endothelial dysfunction in streptozotocininduced 

diabetes in male rats. 

Zhao ZW, Cai W, Lin YL, Lin QF, Jiang Q, Lin Z, Chen LL. 

Source 

Department of Cardiology, Union Hospital, Fujian Medical University, and Fujian 

Institute of Coronary Artery Disease, Fuzhou, PR China. 


The present study was designed to examine whether astaxanthin (ASX, 3,3- 

dihydroxybeta, beta-carotene-4,4-dione, CAS 472-61-7), a dietary antioxidant carotenoid 

that is naturally present in algae, crustaceans, and fish, has a protective effect on 

endothelial dysfunction of aortas in diabetic rats and the possible molecular mechanism 

involved. Male Wistar rats were randomly divided into four groups: control rats, diabetic 

rats, diabetic rats treated with ASX (10 mg/kg/d), and control rats treated with ASX. 

Type 1 diabetes was induced by a single intraperitoneal injection of streptozotocin (STZ; 

60 mg/ kg). STZ-induced diabetes in rats was complicated with excessive oxidative stress 

and endothelial dysfunction, increased serum oxidized low-density lipoprotein (ox-LDL) 

and aortic malondialdehyde (MDA) levels, inhibited endothelium-dependent 

vasorelaxation to acetylcholine (ACh) and unaffected endothelium-dependent 

vasorelaxation to sodium nitroprusside (SNP). Simultaneously, lectin-like oxLDL 

receptor-i (LOX-1) expression was enhanced and endothelial nitric oxide (NO) synthase 

(eNOS) expression was reduced in the aortas of diabetic rats. ASX treatment could 

significantly decrease serum oxLDL and aortic MDA levels, attenuate blunted 

endothelium-dependent vasodilator responses to ACh, upregulate eNOS expression, and 

decrease LOX-1 expression. These results indicated that ASX could ameliorate diabetic 

endothelial dysfunction by inhibiting the ox-LDLLOX-1-eNOS pathway. Treatment with 

ASX might be clinically useful for diabetic complications associated with endothelial 

dysfunction. 


Diabetes 

265

Int J Med Sci. 2011 Feb 9;8(2):126-38. 

High dose astaxanthin lowers blood pressure and 

increases insulin sensitivity in rats: are these effects 

interdependent? 

Preuss HG, Echard B, Yamashita E, Perricone NV. 

Source 

Georgetown University Medical Center, Department of Biochemistry, Washington, DC 

20057, USA. preusshg@georgetown.edu 


The present investigation in Sprague-Dawley rats (SD) was designed to examine effects 

of astaxanthin (Asta) at different doses on elevated blood pressure (BP) and glucoseinsulin 

perturbations produced by heavy sucrose ingestion. We also examined effects of 

Asta on BP during restraint stress. SD were divided into six groups each containing eight 

rats. All SD ate a basic diet of ground regular rat chow with sucrose added at 30% w/w. 

The Control group received only the basic diet containing added sucrose, while the other 

five groups each received the same diet with added test material: captopril, (30 mg/Kg), 

pioglitazone (15.0 mg/Kg), low Asta (25 mg/Kg), medium Asta (50 mg/kg) or high Asta 

(100 mg/Kg). Many tests were carried out to examine the mechanisms behind the effects 

of Asta on BP (serum ACE activity, losartan challenge, and LNAME challenge) and the 

glucose-insulin system (glucose tolerance, HOMA measurement, and insulin challenge). 

In SD, a relatively low dose of Asta decreased SBP, but produced no major changes in 

the glucose-insulin system simulating results from a previous study using Zucker Fatty 

Rats. Increasing the dose of Asta resulted in both a lowering of elevated systolic BP and 

enhanced insulin sensitivity determined by many different estimations. BP lowering was 

consistent with changes in the renin-angiotensin (RAS) and nitric oxide (NO) systems. At 

the examined doses of each, captopril lowered BP in SD without influencing glucoseinsulin 

metabolism, whereas pioglitazone favorably affected glucose-insulin metabolism 

while showing essentially no effects on BP. Accordingly, Asta beneficially affects both 

sucrose-induced elevations of BP and insulin resistance at relatively high doses in SD. 

Also, Asta at higher doses lessens restraint stress, whereas, captopril and pioglitazone did 

not at the doses examined, even though they influenced the BP and glucose-insulin 

systems respectively. 



Diabetes 

266

Int Immunopharmacol. 2011 Jan;11(1):103-9. Epub 2010 Nov 4. 

ROS production in neutrophils from alloxan-induced 

diabetic rats treated in vivo with astaxanthin. 

Marin DP, Bolin AP, Macedo Rde C, Sampaio SC, Otton R. 

Source 

Postgraduate Program, Human Movement Sciences Institute of Physical Activity and 

Sport Sciences, Cruzeiro do Sul University, São Paulo, SP, Brazil, 01506-000. 


BACKGROUND: Astaxanthin (ASTA) is a carotenoid which has powerful 

antioxidant, anti-tumor, anti-diabetic, anti-inflammatory and cardioprotective properties. 

The present study investigated the effect of daily ASTA intake on oxidative stress and the 

functional properties of neutrophils from alloxan-induced diabetic rats. 

METHODS: Neutrophils isolated from ASTA-fed rats (30days, 20mg ASTA/kg of body 

weight - BW) induced to diabetes by alloxan treatment (i.p. 75mg/BW) were assessed by: 

production of superoxide and hydrogen peroxide, nitric oxide, basal calcium release, 

oxidative damage (TBARS and carbonyls content), and activities of major antioxidant 

enzymes. 

RESULTS: Our results show that diabetes promotes a significant oxidative stress in 

neutrophils. The production of superoxide was significantly increased in neutrophils from 

diabetic rats and treatment with ASTA was not effective in reducing superoxide levels. At 

the same time, a reduction in the activity of total superoxide dismutase enzyme was 

observed, which was not restored after treatment with ASTA. At resting conditions, 

neutrophils have a higher basal production of hydrogen peroxide, which is enhanced 

following PMA-stimulation. Treatment with ASTA does not restore values to the basal 

levels. The indicators of oxidative damage to biomolecules showed that diabetic rats 

significantly increased the lipid and protein damage, but this change was reversed after 

treatment with ASTA. 

CONCLUSION: Our results show that diabetes condition promotes a marked 

oxidative stress in neutrophils and treatment with ASTA for 30days at a dose of 20mg/kg 

of BW partially reverses those deleterious effects. 

Copyright Â© 2010 Elsevier B.V. All rights reserved. 


Diabetes 

267

Food Funct. 2011 May;2(5):251-8. Epub 2011 Apr 21. 

Protective actions of microalgae against 

endogenous and exogenous advanced 

glycation endproducts (AGEs) in human 

retinal pigment epithelial cells. 

Sun Z, Liu J, Zeng X, Huangfu J, Jiang Y, Wang M, Chen F. 

Source 

School of Biological Sciences, The University of Hong Kong, Pokfulam Road, Hong 

Kong, P. R. China. 


The formation and accumulation of advanced glycation endproducts (AGEs) is a key 

pathophysiological process involved in various diabetic complications such as diabetic 

retinopathy. In the present study, for the first time, protective effects of three microalgal 

strains, including their extracts and active compounds, against both endogenous and 

exogenous AGEs in cell-based models were investigated. Results showed that in cultured 

human-derived retinal pigment epithelial ARPE-19 cells, the extract of Chlorella 

zofingiensis and its nutritional ingredient astaxanthin exhibited significant inhibitory 

effects on the formation of endogenous N(ε)-carboxymethyllysine (CML), a key AGE 

representative, through the suppression of intracellular oxidative stress. On the other 

hand, extracts of Chlorella zofingiensis, Chlorella protothecoides and Nitzschia laevis as 

well as their nutritional ingredients, namely astaxanthin, lutein and eicosapentaenoic acid 

(EPA), attenuated the deleterious effects induced by exogenous AGEs, such as cell 

proliferation and mRNA upregulation of vascular endothelial growth factor (VEGF) and 

matrix metalloproteinases (MMP)-2, which are critical steps involved in the pathogenesis 

of diabetic retinopathy. These results suggested the positive roles of astaxanthin, lutein 

and EPA in controlling the development of diabetes. These microalgae, therefore, might 

be regarded as beneficial foods and preventive agent choices for patients with diabetic 

retinopathy. 


Diabetes 

268

Ulcers and Gastrointestinal Health 

Eur J Pharmacol. 2005 May 2;514(1):53-9. Epub 2005 Apr 20. 

Protective effect of astaxanthin on naproxen-induced gastric antral 

ulceration in rats. 

Kim JH, Kim YS, Song GG, Park JJ, Chang HI. 


University, Seoul 136-701, South Korea. 

Frequently used for humans as non-steroidal anti-inflammatory drug, naproxen 

has been known to induce ulcerative gastric lesion. The present study investigated 

the in vivo protective effect of astaxanthin isolated from Xanthophyllomyces 

dendrorhous against naproxen-induced gastric antral ulceration in rats. The oral 

administration of astaxanthin (1, 5, and 25 mg/kg of body weight) showed a 

significant protection against naproxen (80 mg/kg of body weight)-induced 

gastric antral ulcer and inhibited elevation of the lipid peroxide level in gastric 

mucosa. In addition, pretreatment of astaxanthin resulted in a significant increase 

in the activities of radical scavenging enzymes such as superoxide dismutase, 

catalase, and glutathione peroxidase. A histologic examination clearly proved that 

the acute gastric mucosal lesion induced by naproxen nearly disappeared after the 

pretreatment of astaxanthin. These results suggest that astaxanthin removes the 

lipid peroxides and free radicals induced by naproxen, and it may offer potential 

remedy of gastric ulceration. 


Ulcers & Gastrointestinal Health 

269

Yao Xue Xue Bao. 2009 May;44(5):558-60. 

[Therapeutic effect of astaxanthin on acetic acid-induced gastric ulcer in 

rats] 


Yang Q, Zhang Z, Zhu X, Ruan H, Fu Y. 

School of Pharmacy, Yanbian University, Yanji 133000, China. 


This study is to investigate therapeutic effect of astaxanthin on acetic acid-induced gastric 

ulcer in rats. Rats were divided into control group, ulcer control group, and astaxanthin 

(5, 10, and 25 mg x kg(-1)) groups at random, 8 rats in each group. After administered for 

10 days consecutively, all the rats were sacrificed. The area of ulcer and the levels of 

MDA, SOD, CAT and GSH-Px in gastric mucosa were measured. Compared with ulcer 

control group, in astaxanthin (5, 10, and 25 mg x kg(-1)) groups, the area of ulcer was 

decreased significantly. Level of MDA decreased while activities of SOD, CAT and 

GSH-Px increased (P < 0.05). Astaxanthin has good therapeutic effect on acetic acidinduced 

gastric ulcer in rats. Eliminating free radical and improving local blood 

circulation of the ulcer may be the mechanism of action. 



270

Eur J Pharmacol. 2008 Aug 20;590(1-3):387-95. Epub 2008 Jun 16. 

Ulcer preventive and antioxidative properties of astaxanthin from 

Haematococcus pluvialis. 

Kamath BS, Srikanta BM, Dharmesh SM, Sarada R, Ravishankar GA. 

Plant Cell Biotechnology Department, Central Food Technological Research 

Institute, Mysore, 570 020, India. 

The anti-ulcer properties of astaxanthin fractions such as total carotenoid and 

astaxanthin esters from Haematococcus pluvialis were evaluated in ethanolinduced 

gastric ulcers in rats. Since oxygen radical release is a pathogenic factor 

of ethanol-induced gastric damage, astaxanthin - a free radical scavenger, was 

investigated as a potential ulcer preventive agent. Astaxanthin fractions - total 

carotenoid and astaxanthin esters were orally administered to experimental rats at 

100, 250 and 500 microg/kg b.w. prior to ulcer induction. Alcian blue binding 

assay indicates that, total carotenoid and astaxanthin esters at 500 microg/kg b.w 

could protect gastric mucin approximately 40% and 67% respectively. Pretreatment 

with astaxanthin esters, also resulted in significant increase in 

antioxidant enzyme levels - catalase, superoxide dismutase, and glutathione 

peroxidase in stomach homogenate. Histopathological examination substantiated 

the protective effect of astaxanthin in pre-treated rats. The increased antioxidant 

potencies such as free radical scavenging activity with an IC(50) of approximately 

8 microg/ml and reducing power abilities (59 x 10(3) U/g) in vitro, reveal that H. 

pluvialis astaxanthin may protect gastric mucosal injury by antioxidative 

mechanism. In addition, approximately 23 fold increased lipoxygenase-inhibitory 

property, in comparison with standard astaxanthin and significant H(+), K(+)- 

ATPase-inhibitory activity of astaxanthin esters, in comparison with known 

proton pump blocking anti-ulcer drug - omeprazole, may envisage the potential 

gastroprotective effect by regulating the gastric mucosal injury and gastric acid 

secretion by the gastric cell during ulcer disease. 




271

Phytomedicine. 2008 Jun;15(6-7):391-9. Epub 2008 May 7. 

Efficacy of the natural antioxidant astaxanthin in the treatment of 

functional dyspepsia in patients with or without Helicobacter pylori 

infection: A prospective, randomized, double blind, and placebocontrolled 

study. 

Kupcinskas L, Lafolie P, Lignell A, Kiudelis G, Jonaitis L, Adamonis K, 

Andersen LP, Wadström T. 

Kaunas University of Medicine, 50009 Kaunas, Lithuania. 

limas.kupcinskas@kmu.lt 

OBJECTIVES: The aim of this study was to evaluate the efficacy of the natural 

antioxidant astaxanthin in functional dyspepsia in different doses and compared 

with placebo. DESIGN: The study was a controlled, prospective, randomized, and 

double blind trial. PARTICIPANTS: Patients with functional dyspepsia, divided 

into three groups with 44 individuals in each group (placebo, 16mg, or 40mg 

astaxanthin, respectively). INTERVENTIONS: Participants were asked to accept 

gastroscopy before treatment, together with questionnaires: GSRS and SF-36. 

Urea breath test (UBT) was done before the treatment. MAIN OUTCOME: The 

primary objective was to test the hypothesis that the antioxidant astaxanthin at 

two doses regimens compared to placebo should ameliorate gastrointestinal 

discomfort measured as GSRS in patients with functional dyspepsia, who were 

either positive or negative for Helicobacter pylori, after 4 weeks of treatment. 

RESULTS: At the end of therapy (week 4) no difference between the three 

treatment groups was observed regarding mean Gastrointestinal Symptom Rating 

Scale (GSRS) scores of abdominal pain, indigestion and reflux syndromes. The 

same results were observed at the end of follow-up. However reduction of reflux 

syndrome before treatment to week 4 was significantly pronounced in the higher 

(40mg) dose compared to the other treatment groups (16mg and placebo, p=0.04). 

CONCLUSION: In general, no curative effect of astaxanthin was found in 

functional dyspepsia patients. Significantly greater reduction of reflux symptoms 

were detected in patients treated with the highest dose of the natural antioxidant 

astaxanthin. The response was more pronounced in H. pylori-infected patients. 




272

FEMS Immunol Med Microbiol. 2007 Jul;50(2):244-8. Epub 2007 May 23. 

Gastric inflammatory markers and interleukins in patients with 

functional dyspepsia treated with astaxanthin. 

Andersen LP, Holck S, Kupcinskas L, Kiudelis G, Jonaitis L, Janciauskas D, 

Permin H, Wadström T. 

Department of Clinical Microbiology, Copenhagen University Hospital, 

Rigshospitalet, Copenhagen, Denmark. lpa@rh.dk 

The chronic active inflammation caused by Helicobacter pylori is dominated by 

neutrophils, macrophages, lymphocytes and plasma cells. Several interleukins are 

involved in the inflammatory process. The aim of this study was to investigate the 

effect of astaxanthin on gastric inflammation in patients with functional 

dyspepsia. Forty-four consecutive patients were included, and biopsies were 

examined for IL-4, IL-6, IL-8, IL-10, interferon-gamma, CD4, CD8, CD14, 

CD19, CD25 and CD30. Patients were randomized: 21 patients were treated with 

40 mg of astaxanthin daily, and 23 patients were treated with a placebo. There 

was a significant decrease in gastric inflammation in H. pylori-positive patients 

from both groups. There were no significant changes in the density of H. pylori or 

in any of the interleukins during or after treatment. There was a significant upregulation 

of CD4 and down-regulation of CD8 in patients with H. pylori treated 

with astaxanthin. Astaxanthin had an effect on the inflammation and on the 

density of H. pylori in mice in a study where the diet could be standardized 

without antioxidants (Bennedsen et al., 1999). These dietary conditions are 

impossible in studies involving humans, and may be due to the minor effect when 

the host have access to antioxidants in their diet. 




273

J Nutr Sci Vitaminol (Tokyo). 2005 Jun;51(3):135-41. 

Effects of astaxanthin and vitamin C on the prevention of gastric 

ulcerations in stressed rats. 

Nishikawa Y, Minenaka Y, Ichimura M, Tatsumi K, Nadamoto T, Urabe K. 

Laboratory of Food Science and Nutrition, College of Nutrition, Koshien 

University, 10-1 Momnijigaoka, Takarazuka, Hyogo, 665-0006, Japan. 

nishizen@gaia.eonet.ne.jp 

Astaxanthin (Asx), one of the carotenoids, is a red pigment in fish and 

Crustaceans, and possesses stronger reduction properties than conventional 

carotenoids, like beta-carotene. However, little is known about the biochemical 

properties and physiological functions of astaxanthin. The effects of astaxanthin 

and vitamin C on stressed rats were studied physiologically and biochemically. 

beta-Carotene and three kinds of astaxanthins, which were extracted from 

Haematococcus and Phaffia, and synthesized chemically, were used in these 

experiments. These rats given astaxanthins or beta-carotene had stress induced on 

the 12th day by immersing the rats in chest-level water at 20 degrees C for 24 h 

after fasting for 24 h. Rats given astaxanthins or beta-carotene prior to stressing 

were appreciably protected against the evolution of gastric ulcerations in relation 

to control rats. Ulcer indexes in particular were smaller with the rat group fed 

astaxanthin extracted from Haematococcus than the other groups. Next, the 

effects of Asx and/or vitamin C on the protection of evolution of gastric ulcer in 

stressed rats were persued by the same methods as described above. The results 

showed that rats given Asx or vitamin C were appreciably protected against the 

evolution of gastric ulcerations in relation to control rats. The effects were more 

intense, especially in rats simultaneously supplied Asx and vitamin C than in rats 

taking either Asx or vitamin C. It was suggested that the simultaneous 

supplementation of food substances with astaxanthin and vitamin C would supply 

enough antioxidants to offset stress-related injuries. 



274

Biosci Biotechnol Biochem. 2005 Jul;69(7):1300-5. 

Suppressive effect of astaxanthin isolated from the 

Xanthophyllomyces dendrorhous mutant on ethanol-induced gastric 

mucosal injury in rats. 

Kim JH, Choi SK, Choi SY, Kim HK, Chang HI. 


University, Seoul 136-701, South Korea. 

Ethanol has been found to induce ulcerative gastric lesion in humans. The present 

study investigated the in vivo protective effect of astaxanthin isolated from the 

Xanthophyllomyces dendrorhous mutant against ethanol-induced gastric mucosal 

injury in rats. The rats were treated with 80% ethanol for 3 d after pretreatment 

with two doses of astaxanthin (5 and 25 mg/kg of body weight respectively) for 3 

d, while the control rats received only 80% ethanol for 3 d. The oral 

administration of astaxanthin (5 and 25 mg/kg of body weight) showed significant 

protection against ethanol-induced gastric lesion and inhibited elevation of the 

lipid peroxide level in gastric mucosa. In addition, pretreatment with astaxanthin 

resulted in a significant increase in the activities of radical scavenging enzymes 

such as superoxide dismutase, catalase, and glutathione peroxidase. A histologic 

examination clearly indicated that the acute gastric mucosal lesion induced by 

ethanol nearly disappeared after pretreatment with astaxanthin. 



275

Clin Microbiol Infect. 2002 Jul;8(7):438-41. 

Effect of antioxidants on the immune response of Helicobacter 

pylori. 

Akyön Y. 

Hacettepe University, School of Medicine, Department of Microbiology and 

Clinical Microbiology, Ankara, Turkey. yakyon@tr.net 

Antioxidants are substances capable of inhibiting oxidation. In chronic diseases, 

inflammatory response cells produce oxygen free radicals. Oxygen free radicals 

cause DNA damage, and this may lead to gene modifications that might be 

carcinogenic. Chronic Helicobacter pylori infection causes the production of 

DNA-damaging free radicals. In recent years, various groups have studied the 

effects of antioxidants, especially on H. pylori-associated gastric cancer. In most 

of the studies, it has been shown that H. pylori infection does affect the level of 

antioxidants measured in the gastric juice, but there are also controversial results. 

Recent experimental studies, both in vivo and in vitro, have shown that vitamin C 

and astaxanthin, a carotenoid, are not only free radical scavengers but also show 

antimicrobial activity against H. pylori. It has been shown that astaxanthin 

changes the immune response to H. pylori by shifting the Th1 response towards a 

Th2 T-cell response. Very few experimental studies support the epidemiologic 

studies, and further studies are needed to describe the effect and the mechanism of 

antioxidants in the H. pylori immune response. 




276

Antimicrob Agents Chemother. 2000 Sep;44(9):2452-7. 

Astaxanthin-rich algal meal and vitamin C inhibit Helicobacter 

pylori infection in BALB/cA mice. 

Wang X, Willén R, Wadström T. 

Department of Infectious Diseases and Medical Microbiology, University of 

Lund, Sweden. 

Helicobacter pylori infection in humans is associated with chronic type B 

gastritis, peptic ulcer disease, and gastric carcinoma. A high intake of carotenoids 

and vitamin C has been proposed to prevent development of gastric malignancies. 

The aim of this study was to explore if the microalga Haematococcus pluvialis 

rich in the carotenoid astaxanthin and vitamin C can inhibit experimental H. 

pylori infection in a BALB/cA mouse model. Six-week-old BALB/cA mice were 

infected with the mouse-passaged H. pylori strain 119/95. At 2 weeks 

postinoculation mice were treated orally once daily for 10 days (i) with different 

doses of algal meal rich in astaxanthin (0.4, 2, and 4 g/kg of body weight, with the 

astaxanthin content at 10, 50, and 100 mg/kg, respectively), (ii) with a control 

meal (algal meal without astaxanthin, 4 g/kg), or (iii) with vitamin C (400 mg/kg). 

Five mice from each group were sacrificed 1 day after the cessation of treatment, 

and the other five animals were sacrificed 10 days after the cessation of treatment. 

Culture of H. pylori and determination of the inflammation score of the gastric 

mucosae were used to determine the outcome of the treatment. Mice treated with 

astaxanthin-rich algal meal or vitamin C showed significantly lower colonization 

levels and lower inflammation scores than those of untreated or control-mealtreated 

animals at 1 day and 10 days after the cessation of treatment. Lipid 

peroxidation was significantly decreased in mice treated with the astaxanthin-rich 

algal meal and vitamin C compared with that of animals not treated or treated 

with the control meal. Both astaxanthin-rich algal meal and vitamin C showed an 

inhibitory effect on H. pylori growth in vitro. In conclusion, antioxidants may be a 

new strategy for treating H. pylori infection in humans. 




277

Immunol Lett. 1999 Dec 1;70(3):185-9. 

Treatment of H. pylori infected mice with antioxidant astaxanthin 

reduces gastric inflammation, bacterial load and modulates cytokine 

release by splenocytes. 

Bennedsen M, Wang X, Willén R, Wadström T, Andersen LP. 

Department of Clinical Microbiology, Rigshospitalet, Copenhagen, Denmark. 

mbe@biobase.dk 

Helicobacter pylori is a gram-negative bacterium affecting about half of the world 

population, causing chronic gastritis type B dominated by activated phagocytes. 

In some patients the disease evolves into gastric ulcer, duodenal ulcer, gastric 

cancer or MALT lymphoma. The pathogenesis is in part caused by the 

immunological response. In mouse models and in human disease, the mucosal 

immune response is characterized by activated phagocytes. Mucosal T- 

lymphocytes are producing IFN-gamma thus increasing mucosal inflammation 

and mucosal damage. A low dietary intake of antioxidants such as carotenoids 

and vitamin C may be an important factor for acquisition of H. pylori by humans. 

Dietary antioxidants may also affect both acquisition of the infection and the 

bacterial load of H. pylori infected mice. Antioxidants, including carotenoids, 

have anti-inflammatory effects. The aim of the present study was to investigate 

whether dietary antoxidant induced modulation of H. pylori in mice affected the 

cytokines produced by H. pylori specific T-cells. We found that treatment of H. 

pylori infected mice with an algal cell extract containing the antioxidant 

astaxanthin reduces bacterial load and gastric inflammation. These changes are 

associated with a shift of the T-lymphocyte response from a predominant Th1- 

response dominated by IFN-gamma to a Th1/Th2-response with IFN-gamma and 

IL-4. To our knowledge, a switch from a Th1-response to a mixed Th1/Th2- 

response during an ongoing infection has not been reported previously. 




278

Antimicrob Agents Chemother. 2000 Sep;44(9):2452-7 

Astaxanthin-rich algal meal and vitamin C inhibit Helicobacter 

pylori infection in BALB/cA mice. 

Wang X, Willen R, Wadstrom T. 

Department of Infectious Diseases and Medical Microbiology, University of 

Lund, Sweden. 

Helicobacter pylori infection in humans is associated with chronic type B 

gastritis, peptic ulcer disease, and gastric carcinoma. A high intake of 

carotenoids and vitamin C has been proposed to prevent development of gastric 

malignancies. The aim of this study was to explore if the microalga 

Haematococcus pluvialis rich in the carotenoid astaxanthin and vitamin C can 

inhibit experimental H. pylori infection in a BALB/cA mouse model. Six-weekold 

BALB/cA mice were infected with the mouse-passaged H. pylori strain 

119/95. At 2 weeks postinoculation mice were treated orally once daily for 10 

days (i) with different doses of algal meal rich in astaxanthin (0.4, 2, and 4 g/kg 

of body weight, with the astaxanthin content at 10, 50, and 100 mg/kg, 

respectively), (ii) with a control meal (algal meal without astaxanthin, 4 g/kg), or 

(iii) with vitamin C (400 mg/kg). Five mice from each group were sacrificed 1 

day after the cessation of treatment, and the other five animals were sacrificed 10 

days after the cessation of treatment. Culture of H. pylori and determination of 

the inflammation score of the gastric mucosae were used to determine the 

outcome of the treatment. Mice treated with astaxanthin-rich algal meal or 

vitamin C showed significantly lower colonization levels and lower 

inflammation scores than those of untreated or control-meal-treated animals at 1 

day and 10 days after the cessation of treatment. Lipid peroxidation was 

significantly decreased in mice treated with the astaxanthin-rich algal meal and 

vitamin C compared with that of animals not treated or treated with the control 

meal. Both astaxanthin-rich algal meal and vitamin C showed an inhibitory 

effect on H. pylori growth in vitro. In conclusion, antioxidants may be a new 

strategy for treating H. pylori infection in humans. 


279

Chem Biol Interact. 2011 Aug 15;193(1):79-87. Epub 2011 May 20. 

Dietary astaxanthin inhibits colitis and colitis-associated colon 

carcinogenesis in mice via modulation of the inflammatory cytokines. 

Yasui Y, Hosokawa M, Mikami N, Miyashita K, Tanaka T. 

Source 

School of Veterinary Medicine, Rakuno Gakuen University, Hokkaido, Japan. y- 

yasui@rakuno.ac.jp 


Astaxanthin (AX) is one of the marine carotenoid pigments, which possess powerful 

biological antioxidant, anti-inflammatory and anti-cancer properties. The purpose of this 

study is to investigate possible inhibitory effect of AX against inflammation-related 

mouse colon carcinogenesis and dextran sulfate sodium (DSS)-induced colitis in male 

ICR mice. We conducted two different experiments. In the first experiment, we evaluated 

the effects of AX at three dose levels, 50, 100 and 200 ppm in diet, on colitis-associated 

colon carcinogenesis induced by azoxymethane (AOM)/DSS in mice. In the second, the 

effects of the AX (100 and 200 ppm) in diet on DSS-induced colitis were determined. We 

found that dietary AX significantly inhibited the occurrence of colonic mucosal ulcers, 

dysplastic crypts, and colonic adenocarcinoma at week 20. AX-feeding suppressed 

expression of inflammatory cytokines, including nuclear factor (NF)-κB, tumor necrosis 

factor (TNF)-α and interleukin (IL)-1β, inhibited proliferation, and induced apoptosis in 

the colonic adenocarcinomas. Feeding with 200 ppm AX, but not 100 ppm, significantly 

inhibited the development of DSS-induced colitis. AX feeding (200 ppm in diet) also 

lowered the protein expression of NF-κB, and the mRNA expression of inflammatory 

cytokines, including IL-1β, IL-6, and cyclooxygenase (COX)-2. Our results suggest that 

the dietary AX suppresses the colitis and colitis-related colon carcinogenesis in mice, 

partly through inhibition of the expression of inflammatory cytokine and proliferation. 

Our findings suggest that AX is one of the candidates for prevention of colitis and 

inflammation-associated colon carcinogenesis in humans. 

Copyright © 2011 Elsevier Ireland Ltd. All rights reserved. 


Ulcers and Gastrointestinal Health 

280

Applications for Athletes 

Int J Sports Med. 2011 Oct 7. [Epub ahead of print] 

Effect of Astaxanthin on Cycling Time 

Trial Performance. 

Earnest CP, Lupo M, White KM, Church TS. 

Source 

Pennington Biomedical Research Center. 


We examined the effect of Astaxanthin (AST) on substrate metabolism and cycling time 

trial (TT) performance by randomly assigning 21 competitive cyclists to 28d of 

encapsulated AST (4 mg/d) or placebo (PLA) supplementation. Testing included a 

VO2max test and on a separate day a 2 h constant intensity pre-exhaustion ride, after a 

10 h fast, at 5% below VO2max stimulated onset of 4 mmol/L lactic acid followed 5 min 

later by a 20 km TT. Analysis included ANOVA and post-hoc testing. Data are Mean 

(SD) and (95% CI) when expressed as change (pre vs. post). Fourteen participants 

successfully completed the trial. Overall, we observed significant improvements in 20 km 

TT performance in the AST group (n=7; -121 s; 95%CI, -185, -53), but not the PLA 

(n=7; -19 s; 95%CI, -84, 45). The AST group was significantly different vs. PLA 

(P

Carotenoid Science, Vol.13, 2008 ISSN 1880-5671 

Dietary Supplementation with Astaxanthin-Rich Algal Meal 

Improves Strength Endurance – A Double Blind Placebo 

Controlled Study on Male Students 

Curt L. Malmsten and Åke Lignell 

The present study was designed to investigate the effect of dietary 

supplementation with astaxanthin on physical performance. Forty healthy 

paramedic students were recruited for this test in a double blind placebo 

controlled study. In this study, we used algal meal (AstaREAL® biomass) as 

astaxanthin supplementation. Twenty of the subjects received capsules filled with 

algal meal to provide 4 mg astaxanthin per capsule, whereas the other twenty 

received placebo capsules for six months. The physical parameters monitored 

were fitness, strength/endurance and strength/explosivity by standardized 

exercises. Before starting the dietary supplementation, base values for each of the 

subjects were obtained. At the end of the six month period of dietary 

supplementation, the average number of knee bendings (squats) increased by 

27.05 (from 49.32 to 76.37) for subjects having received astaxanthin and by 9.0 

(from 46.06 to 55.06) for the placebo subjects. Hence, the increase in the 

astaxanthin supplemented group was three times higher than that of the placebo 

group (P=0.047). None of the other parameters monitored differed significantly 

between the groups at the end of the study period. Based on this findings, it 

suggested that supplementation of astaxanthin is effective for the improvement of 

strength endurance that may lead to sports performance. 

Applications for Athletes: Strength & Endurance 

282

Biochem Biophys Res Commun. 2008 Feb 22;366(4):892-7. Epub 2007 Dec 17. 

Astaxanthin improves muscle lipid metabolism in exercise via 

inhibitory effect of oxidative CPT I modification. 

Aoi W, Naito Y, Takanami Y, Ishii T, Kawai Y, Akagiri S, Kato Y, Osawa T, 

Yoshikawa T. 

Department of Inflammation and Immunology, Graduate School of Medical 

Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan. 

Intracellular redox balance may affect nutrient metabolism in skeletal muscle. 

Astaxanthin, a carotenoid contained in various natural foods, exerts high 

antioxidative capacity in the skeletal muscles. The present study investigated the 

effect of astaxanthin on muscle lipid metabolism in exercise. ICR mice (8 weeks 

old) were divided into four different groups: sedentary, sedentary treated with 

astaxanthin, running exercise, and exercise treated with astaxanthin. After 4 

weeks of treatment, exercise groups performed treadmill running. Astaxanthin 

increased fat utilization during exercise compared with mice on a normal diet with 

prolongation of the running time to exhaustion. Colocalization of fatty acid 

translocase with carnitine palmitoyltransferase I (CPT I) in skeletal muscle was 

increased by astaxanthin. We also found that hexanoyl-lysine modification of 

CPT I was increased by exercise, while astaxanthin prevented this increase. In 

additional experiment, we found that astaxanthin treatment accelerated the 

decrease of body fat accumulation with exercise training. Our results suggested 

that astaxanthin promoted lipid metabolism rather than glucose utilization during 

exercise via CPT I activation, which led to improvement of endurance and 

efficient reduction of adipose tissue with training. 


 



Applications for Athletes: Endurance 

283

Biol Pharm Bull. 2006 Oct;29(10):2106-10. 

Effects of astaxanthin supplementation on exercise-induced fatigue 

in mice. 


Laboratory of Nutraceuticals and Functional Foods Science, Graduate School of 



The present study was designed to determine the effect of astaxanthin on 

endurance capacity in male mice aged 4 weeks. Mice were given orally either 

vehicle or astaxanthin (1.2, 6, or 30 mg/kg body weight) by stomach intubation 

for 5 weeks. The astaxanthin group showed a significant increase in swimming 

time to exhaustion as compared to the control group. Blood lactate concentration 

in the astaxanthin groups was significantly lower than in the control group. In the 

control group, plasma non-esterfied fatty acid (NEFA) and plasma glucose were 

decreased by swimming exercise, but in the astaxanthin group, NEFA and plasma 

glucose were significantly higher than in the control group. Astaxanthin treatment 

also significantly decreased fat accumulation. These results suggest that 

improvement in swimming endurance by the administration of astaxanthin is 

caused by an increase in utilization of fatty acids as an energy source. 



284

Antioxid Redox Signal. 2003 Feb;5(1):139-44. 

Astaxanthin limits exercise-induced skeletal and cardiac muscle 

damage in mice. 

Aoi W, Naito Y, Sakuma K, Kuchide M, Tokuda H, Maoka T, Toyokuni S, 

Oka S, Yasuhara M, Yoshikawa T. 

Department of Medicine, Kyoto Prefectural University of Medicine, Kyoto, 602- 

0841, Japan. waoi@basic.kpu-m.ac.jp 

Dietary antioxidants may attenuate oxidative damage from strenuous exercise in 

various tissues. Beneficial effects of the antioxidant astaxanthin have been 

demonstrated in vitro, but not yet in vivo. We investigated the effect of dietary 

supplementation with astaxanthin on oxidative damage induced by strenuous 

exercise in mouse gastrocnemius and heart. C57BL/6 mice (7 weeks old) were 

divided into groups: rested control, intense exercise, and exercise with astaxanthin 

supplementation. After 3 weeks of exercise acclimation, both exercise groups ran 

on a treadmill at 28 m/min until exhaustion. Exercise-increased 4-hydroxy-2- 

nonenal-modified protein and 8-hydroxy-2'-deoxyguanosine in gastrocnemius and 

heart were blunted in the astaxanthin group. Increases in plasma creatine kinase 

activity, and in myeloperoxidase activity in gastrocnemius and heart, also were 

lessened by astaxanthin. Astaxanthin showed accumulation in gastrocnemius and 

heart from the 3 week supplementation. Astaxanthin can attenuate exerciseinduced 

damage in mouse skeletal muscle and heart, including an associated 

neutrophil infiltration that induces further damage. 


Applications for Athletes: Prevention of Skeletal and Cardiac Muscle Damage 

285


Sports Performance Benefits from Taking Natural Astaxanthin 

Characterized by Visual Acuity and Muscle Fatigue Improvement in 

Humans 

SAWAKI KEISUKE; YOSHIGI HIROSHI; AOKI KAZUHIRO; KOIKAWA 

NATSUE; AZUMANE AKITO; KANEKO KESATOKI; YAMAGUCHI 

MASAHIRO 

The effects of astaxanthin on visual acuity and muscle fatigue were studied. 

Astaxanthin (3,3'-Dihydroxy-.BETA.,.BETA.-carotene-4,4'-dione) is a red 

pigment found in salmon and krill and has strong antioxidant properties. In the 

two supplementation studies, astaxanthin extracted from algae (Haematococcus 

pluvialis) was used. Four visual acuity parameters were examined in experiment 

A in 18 healthy adult male volunteers that were equally divided into two groups 

(treatment and control). The measured parameters were deep vision, critical 

flicker fusion, static and kinetic visual acuity before and after supplementation. A 

second investigation (experiment B) involved 16 adult male volunteers to 

establish the effect of astaxanthin supplementation on the build up of lactic acid 

before and after running 1200 metres. In both experiments, the treated groups 

ingested an astaxanthin capsule per day for 4 weeks (6mg astaxanthin per day) 

and the control groups received a placebo capsule. Results: In experiment A, the 

deep vision and the critical flicker fusion of the treated groups were significantly 

improved compared to the control group. No effects of treated group were 

observed on static and kinetic visual acuity. In experiment B, serum lactic acid 

concentration at 2 minutes after activity (1,200m running) of the treatment group 

was significantly lower than that of the control one. No other effects related to 

supplementation of astaxanthin on serum biological and hematological 

examinations were observed. Based on these preliminary findings, it suggested 

that supplementation of astaxanthin is effective for the improvement of visual 

acuity and muscle fatigue that may lead to sports performance benefits. 

Applications for Athletes: Visual Acuity and Muscle Fatigue 

286

Mera Pharmaceuticals, Inc. Review presented at the 1 st Congress of the International Society for 

Applied Phycology/9 th International Conference on Applied Phycology, May 2002, Almeria, 

Spain. 

Haematococcus astaxanthin: health and nutrition applications: 

Exercise survey with 88% reporting improvement 

Guerin, M, Huntley, M, Olaizola, M. 

“In March 2001, a health survey looked at the various positive effects of Astaxanthin on 

exercise. The survey involved 247 between the ages of 20 and 87 years. 146 of those 

taking part reported problems with muscle and joint soreness. When taking Astaxanthin, 

88% of participants reported improvement. In all cases, the more exercise an individual 

did, the more benefit was experienced.” 

Applications for Athletes: Muscle and Joint Soreness 

287

PhysioL Chern. Phys. & M«l. NMR. (1990) 22:27-31:\ 

Inhibition of Oxidative Injury of Biological Membranes by 

Astaxanthin 

Michi Kurashige, Eiji Okimasu, Masayasu Inoue and Kozo Utsumi 

The value of astaxanthin, a carotenoid pigment, in the treatment of oxidative 

injury is assessed. Astaxanthin protects the mitochondria or vitamin E-deficient 

rats from damage by Fe2+ catalyzed lipid peroxidation both in vim and in vitro. 

The inhibitory effect of astaxanthin on mitochondrial lipid peroxidation is 

stronger than that of a-tocopherol. Thin layer chromatographic analysis shows 

that the change in phospholipid components of erythrocytes from vitamin E- 

deficient rats induced by Fe²+ and Fe³+ -xanthine/xanthine oxidase system was 

significantly suppressed by astaxanthin. Carrageenan-induces inflammation of 

the paw is also significantly inhibited by administration of astaxanthin. These 

data indicate that astaxanthin functions as a potent antioxidant both in vivo and in 

vitro. 

Applications for Athletes: Mitochondria (Energy Producing Part of Cells) 

288

J Nutr Biochem. 2009 May 6. [Epub ahead of print] 

Astaxanthin protects mitochondrial redox state and functional integrity 

against oxidative stress. 

Wolf AM, Asoh S, Hiranuma H, Ohsawa I, Iio K, Satou A, Ishikura M, Ohta S. 

Department of Biochemistry and Cell Biology, Institute of Development and Aging 

Sciences, Nippon Medical School, Nakahara-ku, Kawasaki, Kanagawa 211-8533, Japan. 

Mitochondria combine the production of energy with an efficient chain of reductionoxidation 

(redox) reactions but also with the unavoidable production of reactive oxygen 

species. Oxidative stress leading to mitochondrial dysfunction is a critical factor in many 

diseases, such as cancer and neurodegenerative and lifestyle-related diseases. Effective 

antioxidants thus offer great therapeutic and preventive promise. Investigating the 

efficacy of antioxidants, we found that a carotenoid, astaxanthin (AX), decreased 

physiologically occurring oxidative stress and protected cultured cells against strong 

oxidative stress induced with a respiratory inhibitor. Moreover, AX improved 

maintenance of a high mitochondrial membrane potential and stimulated respiration. 

Investigating how AX stimulates and interacts with mitochondria, a redox-sensitive 

fluorescent protein (roGFP1) was stably expressed in the cytosol and mitochondrial 

matrix to measure the redox state in the respective compartments. AX at nanomolar 


Additionally, AX improved the ability of mitochondria to remain in a reduced state under 

oxidative challenge. Taken together, these results suggest that AX is effective in 

improving mitochondrial function through retaining mitochondria in the reduced state. 


Applications for Athletes: Mitochondria (Energy Producing Part of Cells) 

289

Human Performance Laboratories, The University of Memphis, 

Memphis, TN, USA 38152 

ASTAXANTHIN SUPPLEMENTATION 

A.C. Fry, B.K. Schilling, L.Z.F. Chiu, N. Hori, and L.W. Weiss, FACSM. 


PURPOSE: To determine the effects of astaxanthin anti-oxidant supplementation 

as a counter-measure for delayed onset muscular soreness (DOMS) in currently 

trained individuals, nine weight trained males (X+SE: age=25.1+1.6 yrs., 

hgt=1.79+0.02 m, wgt=86.8+4.4 kg) participated in this study. METHODS: All 

subjects provided muscle biopsy samples from the vastus lateralis m. prior to 

inducing DOMS in the knee extensor mm. (10 sets x 7-10 reps, 85% eccentric 1 

RM). The subjects ingested either 4 mg.d-1 of astaxanthin (Suppl; n=4) or a 

placebo (Con; n=5) for a 3 week loading phase prior to the DOMS-inducing 

protocol, and during a 12 d recovery phase. Perceptions of DOMS at 48 hrs posteccentric 

exercise were quantified by muscle soreness ratings (0-10 Likert scale). 

Muscle fiber characteristics were determined via mATPase histochemistry and 

digital imaging to determine % cross-sectional areas of the major fiber types (I, 

IIA, IIAB/B). Due to small numbers of IIB fibers in some subjects, IIAB hybrid 

fibers were included in this fiber type population. Simple regression was used to 

determine relationships between fiber characteristics and perceptions of soreness. 

RESULTS: No differences in perceptions of soreness between the Suppl or Con 

groups were observed (p>0.05), with all subjects exhibiting a mean score of >5. 

Percent fiber type areas were similar (p>0.05) for both groups (type I, 

Suppl=47.6+8.9%, Con=41.3+2.7%; type IIA, Suppl=44.3+5.6%, 

Con=53.0+2.8%; type IIAB/B, Suppl=8.2+3.6%, Con=5.7+1.6%). However, 48 

hrs after the DOMS-inducing session, perceptions of soreness for the Suppl group 

were positively related to % area type I (r=0.90), and negatively related to % area 

types IIA (r=-0.80) and IIAB/B (r=-0.99). A distinctly different correlational 

pattern was observed for the Con group (% type I area, r=-0.58; % type IIA area, 

r=0.32; % type IIAB/B area, r=0.40). CONCLUSIONS: Collectively, these 

preliminary data suggest that astaxanthin supplementation may preferentially 

attenuate perceptions of DOMS in weight trained men with a high % area for fiber 

types IIA & AB/B. 

Applications for Athletes: Delayed Onset Muscle Soreness 

290

Hiro to Kyuyo no Kagaku VOL.18;NO.1;PAGE.35-46(2003) 

Effects of Astaxanthin on Recovery from Whole Fatigue with Three 

Stepwise Exercises 

NAGATA AKIRA; TAJIMA TAEKO; HAMAMATSU HOZUMI 

This study was designed to evaluate the effects of astaxanthin (A) ingestion upon 

recovery from whole fatigue, that were generated by progressive loads of three 

stepwise exercise-30%HRmax, 50%HRmax, and 70%HRmax. Nineteen healthy 

volunteers were randomized into two groups: Group A (10 subjects) received oral 

astaxanthin capsule (5mg) daily for two weeks, while Group C (9 subjects) 

ingested oral placebo (C) capsule (5mg) with the double blind method. After a 

month from this ingestion, another capsules were taken again with cross-over 

system for the same subjects respectively. Comparative detections were practiced 

to estimate with effectiveness of A ingestion upon changing ratios between two 

groups. Significant difference between A and C groups were obtained to inhibit 

the increase of respiratory-circulatory function from expired gases analysis. 

Additionally sympathetic nervous activities (LF/HF ratio) during exercise and 

parasympathetic nervous activities (HF/TF 100) during recovery were observed to 

significant increase. Otherwise, blood serum concentration of LDL cholesterols 

showed significant decrease, while concentration of creatine phosphokinase had 

increased to higher level than that of C ingestion, significantly. Then, findings of 

the present study indicated that with astaxanthin ingestion for human, respiratorycirculation 

ability and activities of sympathetic nervous system were augmented 

to make efficient metabolism during exercise load. Those anti-fatigue and antioxidative 

function might be promoted for human to make recovery ability from 

the whole fatigue generated by exercise stress. 

Applications for Athletes: Recovery 

291

Japanese Journal of Physical Fitness and Sports Medicine. Vol. 54, No. 6, pg 466. December 

2005. 

Effect of Astaxanthin on Muscular Atrophy 

Tateo Sugiura, Yoshiharu Iida, Hisashi Naito, Daijiro Ohmori, Katsumasa Goto, 

Toshitada Yoshioka 

Objective: Patients wearing casts or other devices that hinder mobility are reported to 

have muscular atrophy. It is commonly thought that the cause is from reactive oxygen 

species (ROS). The use of Vitamin E, along with other antioxidants, prevents ROS from 

causing muscular atrophy that arises from lack of movement; however there has been 

conflicting reports on its effectiveness, varying from some claiming that it works and 

others that it does not. 

Results and Analysis: Groups that were administered Ax had significantly less muscle 

atrophy than those in the Control group (p

Long term dietary antioxidant intakes attenuate sarcopenia 

Tsubasa SHIBAGUCHll, Talmo SUGIURAl, Tsukasa FURUMOTOl, Koshiro 

I~OUEI, Yoshiharu TlDA2,Hieeebl ~AIToa, Kaeeumaea GOTO', Daijiro 

OHMORI~, 'Ibshitadu YOSmOK.,V 

Department of Exercise and Health Sciences, Faculty of Education, Yamaguchi 

University. Yamaguchi 753-8513, Japan, 2'fbyo Koso Kagaku Co. Ltd. Ureveeu, Chiba, 

279-00U. Japan, Department of Exercise Physiology , School of Health and Sports 

Science, Junteodo University, Inba, Cbibe 270-1695, Japen, Laboratory Physiology, 

Toyobashi SOZO University, Toyohaehi 440-8511, 6 Department of Chemistry, School 

of Medicine, Juotendo University Japan, 6Hiroaaki Oakum University, Hircsaki, Aomori 

036-8577, Japan 

Japanese Journal of Physical Fitness and Sports Medicine. 2008, 57:541-52 

Oxidative stress is thought to be one of significant contributing factors to agerelated 

sarccpenia. We tested the hypothesis that the long term dietary antioxidant 

(astaxanthin intakes attenuate sarcopenie. Wistar strain male rats, aged 45 weeks 

old, were given either control (Cont) or astaxanthin feed (0.004%, Ax) for 1 year. 

'The soleus muscle weights and muscle weigh-to-body weight ratios in Ax group 

were significantly heavier than in Cont group, but tibialis anterior muscle mass 

remained similar between the two dietary groups. The level of ubiquitinated 

proteins was significantly lower in soleus muscles of Ax group, but not in tibialis 

anterior muscles when compared with Cont group. Tibialis anterior levels of 

cathepsin Land caepase-S were tended to be lower in Ax group than in Cont 

group, especially significant differences observed in cathepsin L, whereas no 

differences between Cont and Ax were observed in soleus tbcae levels. There 

woro no effects of Ax supplementation on calpaia 1 and 2, UBC3B, CulZn SOD 

and nitrotyrosine levels in both soleus and tibialis anterior muscles. OUT data 

suggest that the long term dietary astaxanthin intakes attenuate the age related 

muscle atrophy, due in part, to reductions in oxidative stress and ubiquitination of 

myofibrillar protein in slow soleus muscles, but not in fast tibialis anterior 

muscles. 

Applications for Athletes: Sarcopenia 

293

Title;Effects of Astaxanthin 

Supplementation on 

Exercise-Induced Fatigue 

in Mice 

Author; IKEUCHI MAYUMI (Graduate School of Marine Sci. and Technol., 

Tokyo Univ. of Marine Sci. and Technol.) KOYAMA TOMOYUKI (Graduate 

School of Marine Sci. and Technol., Tokyo Univ. of Marine Sci. and Technol.) 

TAKAHASHI JIRO (Fuji Chemical Industry Co., Ltd.) YAZAWA 

KAZUNAGA (Graduate School of Marine Sci. and Technol., Tokyo Univ. of Marine 

Sci. and Technol.) 

Journal Title;Biol Pharm Bull 

Journal Code:S0989A 

ISSN:0918-6158 

VOL.29;NO.10;PAGE.2106-2110 (J-STAGE)(2006) 

Figure&Table&Reference;FIG.6, REF.22 

Pub. Country;Japan 

Language;English 

Abstract;The present study was designed to determine the effect of astaxanthin on 

endurance capacity in male mice aged 4 weeks. Mice were given orally either vehicle 

or astaxanthin (1.2, 6, or 30 mg/kg body weight) by stomach intubation for 5 weeks. 

The astaxanthin group showed a significant increase in swimming time to exhaustion 

as compared to the control group. Blood lactate concentration in the astaxanthin 

groups was significantly lower than in the control group. In the control group, plasma 

non-esterfied fatty acid (NEFA) and plasma glucose were decreased by swimming 

exercise, but in the astaxanthin group, NEFA and plasma glucose were significantly 

higher than in the control group. Astaxanthin treatment also significantly decreased fat 

accumulation. These results suggest that improvement in swimming endurance by the 

administration of astaxanthin is caused by an increase in utilization of fatty acids as an 

energy source. 


294

Effects of Astaxanthin Ingestion on Exercise-Induced 

Physiological Changes 

Authors: Taeko Tajima, Akira Nagata. Health and Behavior Sciences.,3(1):5- 

10(2004). 


The purpose of this study was to evaluate the effects of astaxantin (ACT) ingestion on 

exercise-induced physiological functions. In this experiment we planned to investigate 

the autonomic nervous system (ANS) and the respiratory metabolism during different 

exercise intensities in subjects taking astaxantin and those taking placebo. The design of 

this experiment was a double-blind crossover study. 

Eighteen male volunteers (35.8 + 4.51 years of age) took ACT or placebo (CON) capsule 

daily for two weeks. Exercise stress tests were done before and after the ingestion 

period. The exercise load was in the form of running exercise on a treadmill at intensities 

of 30%, 50% and 70% of maximum heart rate (HRmax). Heart rate variability (HRV), 

expired gases analysis and blood biochemical parameters were measured. Sympathetic 

nervous activity (SNA) and parasympathetic nervous activity (PNA) were estimated from 

the pattern of power density in three frequency ranges on the power spectrum. 

During the exercise at an intensity of 70% HRmax, CVRR and HF/TF increased 

significantly (p

Additional Areas of Research 

J. Nat. Prod. 2006, 69, 443-449 

Astaxanthin, a Carotenoid with Potential in Human Health and 

Nutrition 

Ghazi Hussein,*,†,‡ Ushio Sankawa,† Hirozo Goto,§ Kinzo Matsumoto,‡ and 

Hiroshi Watanabe‡ 

Astaxanthin (1), a red-orange carotenoid pigment, is a powerful biological 

antioxidant that occurs naturally in a wide variety of living organisms. The potent 

antioxidant property of 1 has been implicated in its various biological activities 

demonstrated in both experimental animals and clinical studies. Compound 1 has 

considerable potential and promising applications in human health and nutrition. 

In this review, the recent scientific literature (from 2002 to 2005) is covered on 

the most significant activities of 1, including its antioxidative and antiinflammatory 

properties, its effects on cancer, diabetes, the immune system, and 

ocular health, and other related aspects. We also discuss the green microalga 

Haematococcus pluVialis, the richest source of natural 1, and its utilization in the 

promotion of human health, including the antihypertensive and neuroprotective 

potentials of 1, emphasizing our experimental data on the effects of dietary 

astaxanthin on blood pressure, stroke, and vascular dementia in animal models, is 

described. 

Additional Areas of Research: General Health 

296

Recenti Prog Med. 2010 Apr;101(4):145-56. 

[Omega-3 fatty acids and astaxanthin in health and disease. Recent 

knowledges] 

[Article in Italian] 

Testino G, Ancarani O, Sumberaz A. 

Dipartimento Medicina Specialistica, Azienda Ospedaliera-Universitaria Ospedale San 

Martino, Genova. testinogia@tiscalinet.it 

Erratum in: 

 

Recenti Prog Med. 2010 May;101(5):180. 


At present, medicine is aimed to the treatment of lesions. Instead, it would be right to 

develop the maintenance of normal health. A number of authorities have recently 

recommended increases in intake of omega-3 fatty acids and astaxanthin for the health of 

general population. Omega-3 are necessary to provide the optimal function of cellular 

membrane in health and in disease states. It is well known how at least two servings of 

fish a week, or dietary supplementation of fatty acids omega-3, should be taken to obtain 

the health benefits of this essential nutrient. Astaxanthin is a powerful biological 

antioxidant. This property has been implicated in its various biological activities 

demonstrated in both experimental animals and clinical studies. For the recent evidence 

of the contemporary presence of omega-3 and astaxanthin in oil of Wild Pacific Salmon 

Sockeye, a review has been effected for the evaluation of a possible role of such 

association for the health promotion. 



297

Critical Reviews in Food Science and Nutrition, 46:185–196 (2006) ISSN: 1040-8398 

Astaxanthin: A Review of its Chemistry and Applications 

HIGUERA-CIAPARA, L. FE´ LIX-VALENZUELA, and F. M. GOYCOOLEA 














recent and has mostly been performed in vitro or at the 

pre-clinical level with humans. This paper reviews the current available evidence 

regarding astaxanthin chemistry and its potential beneficial effects in humans. 


298

Trends Biotechnol. 2003 May;21(5):210-6. 

Haematococcus astaxanthin: applications for human health and 

nutrition 

Martin Guerin, Mark E, Huntley and Miguel Olaizola 

The carotenoid pigment astaxanthin has important applications in the 

nutraceutical, cosmetics, food and feed industries. Haematococcus pluvialis is the 

richest source of natural astaxanthin and is now cultivated at industrial scale. 

Astaxanthin is a strong coloring agent and a potent antioxidant - its strong 

antioxidant activity points to its potential to target several health conditions. This 

article covers the antioxidant, UV-light protection, anti-inflammatory and other 

properties of astaxanthin and its possible role in many human health problems. 

The research reviewed supports the assumption that protecting body tissues from 

oxidative damage with daily ingestion of natural astaxanthin might be a practical 

and beneficial strategy in health management. 


299

Natural Healing Track August 2000 

ASTAXANTHIN 

Continuing Education Module 

by Timothy J. Maher, Ph.D. 

Goal: 

The goal of this module is to introduce the reader to the carotenoid astaxanthin 

and examine its antioxidant actions especially as it relates to potential therapeutic 

approaches in addressing cardiovascular disease, neurodegenerative disease, 

cancer, immune function status and visual health. 

Objectives: 

Following successful completion of this module, the participant will be able to: 

• describe the unique antioxidant features of the carotenoid astaxanthin; 

• list the sources in nature and the functions of astaxanthin in animals that produce 

and consume astaxanthin; 

• explain findings of recent research that describe the effects of astaxanthin in 

cardiovascular disease, neurodegenerative 

disease, visual health, cancer and immune system function; 

• describe the pharmacokinetics of astaxanthin and list its potential side effects. 


300

Copyright © 2002 by Mera Pharmaceuticals, Inc. All rights reserved. 

Haematococcus astaxanthin: health and nutritional applications 

Martin Guerin, Mark E. Huntley, Miguel Olaizola 

Mera Pharmaceuticals, Inc. 

This review was presented at the 1st Congress of the International Society for 

Applied Phycology/9 th International Conference on Applied Phycology May 26- 

30, 2002, Almeria, Spain 


Astaxanthin, a carotenoid pigment, has important applications in the nutraceutical, 

cosmetics, food and feed industries. Haematococcus pluvialis is the richest source 

of natural astaxanthin and is now cultivated at industrial scale. Astaxanthin is a 

strong coloring agent and a potent antioxidant. Astaxanthin.s strong antioxidant 

activity points to its potential to target a number of health conditions. Here we 

review the scientific literature on antioxidant, UV-light protection, and antiinflammatory 

properties of astaxanthin, and its possible role in cellular health, 

cancer, immunology, liver function, heart health, eye health, central nervous 

system health, and other human health concerns. We also report results of a 

survey among users of a commercially available astaxanthin product 

from 20 to 87 years old. The reported effects of AstaFa 

conform to expectations of astaxanthin activity in chemical and animal models. 

Eighty eight percent of respondents reporting that they suffer from sore muscles 

or joints, observed a reduction in soreness or pain. Similarly, over 80% of those 

reporting back pain and symptoms from osteoarthritis or rheumatoid arthritis 

reported an improvement from 

astaxanthin supplementation. Astaxanthin supplementation was also reported to 

improve symptoms of asthma and enlarged prostate. All of these conditions have 

an inflammation component which is closely tied to oxidative damage. These 

results support the assumption that protecting body tissues from oxidative damage 

with daily ingestion of natural astaxanthin may be a practical and beneficial 

strategy in health management. 


301

Biosci Biotechnol Biochem. 2007 Apr;71(4):893-9. Epub 2007 Apr 7. 

Effects of astaxanthin in obese mice fed a high-fat diet. 


Laboratory of Nertraceuticals and Functional Foods Science, Graduate School of 




living organisms. We investigated the effects of astaxanthin supplementation in 

obese mice fed a high-fat diet. Astaxanthin inhibited the increases in body weight 

and weight of adipose tissue that result from feeding a high-fat diet. In addition, 

astaxanthin reduced liver weight, liver triglyceride, plasma triglyceride, and total 

cholesterol. These results suggest that astaxanthin might be of value in reducing 

the likelihood of obesity and metabolic syndrome in affluent societies. 


Additional Areas of Research: Weight Loss 

302

Methods Find Exp Clin Pharmacol. 2001 Mar;23(2):79-84. 

Effect of astaxanthin on the hepatotoxicity, lipid peroxidation and 

antioxidative enzymes in the liver of CCl4-treated rats. 

Kang JO, Kim SJ, Kim H. 

Department of Food and Nutrition, College of Human Ecology, Seoul National 

University, Korea. 

Astaxanthin is one of many carotenoids present in marine animals, vegetables and 

fruits. Since carotenoids are known to have antioxidant properties, we tested to 

determine if astaxanthin could have protective effects in the CCl4-treated rat liver 

by activating the antioxidant system. Astaxanthin blocked the increase of 

glutamate-oxalacetate transaminase (GOT) and glutamate-pyruvate transaminase 

(GTP) activity and thiobarbituric acid reactive substances (TBARS) in response to 

carbon tetrachloride (CCl4), while causing an increase in glutathione (GSH) 

levels and superoxide dismutase (SOD) activities in the CCl4-treated rat liver. 

These results suggest that astaxanthin protects liver damage induced by CCl4 by 

inhibiting lipid peroxidation and stimulating the cellular antioxidant system. 


 



Additional Areas of Research: Hepatoprotective (Liver) 

303

Xenobiotica. 1996 Sep;26(9):909-19. 

beta-Apo-8'-carotenal, but not beta-carotene, is a strong inducer of 

liver cytochromes P4501A1 and 1A2 in rat. 

Gradelet S, Leclerc J, Siess MH, Astorg PO. 

Unité de Toxicologie Nutritionnelle, Institut National de la Recherche 

Agronomique, Dijon, France. 

1. The catalytic activities of several phase I and II xenobiotic-metabolizing 

enzymes and their immunochemical detection have been investigated in liver 

microsomes and cytosol of the male rat, which had been fed for 15 days with diets 

containing 300 mg/kg beta-carotene isomers (all-trans beta-carotene or betacarotene 

from Dunaliella salina rich in 9-cis isomer or isomerized beta-carotene), 

or apocarotenoids as beta-apo-8'-carotenal, ethyl beta-apo-8'-carotenoate and 

citranaxanthin. 2. Beta-carotene, either all-trans or containing cis isomers, did not 

induce any significant change in the measured activities. By contrast, beta-apo-8'- 

carotenal increased the liver content of cytochrome P450, the activity of NADHand 

NADPH-cytochrome c reductase, and strongly increased some cytochrome 

P450-dependent activities, particularly ethoxyresorufin O-deethylase (x158), 

methoxyresorufin O-demethylase (x22), pentoxy- and benzoxyresorufin O- 

dealkylases, but did not affect erythromycin N-demethylase nor 

nitrosodimethylamine N-demethylase activities. Phase II p-nitrophenol- and 4- 

hydroxy- biphenyl-uridine diphosphoglucuronosyl transferase activities were also 

increased by beta-apo-8'carotenal. Western blots of microsomal proteins clearly 

showed the induction of CYP1A1 and 1A2 by beta-apo-8'-carotenal. This 

induction profile resembles that produced by two other carotenoids: canthaxanthin 

and astaxanthin. Ethyl beta-apo-8'-carotenoate and citranaxanthin showed similar 

effects to beta-apo-8'-carotenal but of less intensity. 3. Three carotenoids: betaapo-8'-carotenal, 

canthaxanthin and astaxanthin, are inducers of CYP1A1 and 

1A2 in the rat. These carotenoids form a new class of inducers of CYP1A, 

structurally very different from the classical inducers as 3-methylcholanthrene, 

beta-naphtoflavone or dioxin. 


 




304

Toxicology. 2010 Jan 12;267(1-3):147-53. Epub 2009 Nov 10. 

Effect of astaxanthin on hepatocellular injury following ischemia/reperfusion. 

Curek GD, Cort A, Yucel G, Demir N, Ozturk S, Elpek GO, Savas B, Aslan M. 

Department of Biochemistry, Akdeniz University Medical School, Antalya 07070, 

Turkey. 


This study investigated the effect of astaxanthin (ASX; 3,3-dihydroxybeta, beta-carotene-4,4- 

dione), a water-dispersible synthetic carotenoid, on liver ischemia-reperfusion (IR) injury. 

Astaxanthin (5 mg/kg/day) or olive oil was administered to rats via intragastric intubation for 14 

consecutive days before the induction of hepatic IR. On the 15th day, blood vessels supplying the 

median and left lateral hepatic lobes were occluded with an arterial clamp for 60 min, followed 

by 60 min reperfusion. At the end of the experimental period, blood samples were obtained from 

the right ventricule to determine plasma alanine aminotransferase (ALT) and xanthine oxidase 

(XO) activities and animals were sacrificed to obtain samples of nonischemic and postischemic 

liver tissue. The effects of ASX on IR injury were evaluated by assessing hepatic ultrastructure 

via transmission electron microscopy and by histopathological scoring. Hepatic conversion of 

xanthine dehygrogenase (XDH) to XO, total GSH and protein carbonyl levels were also measured 

as markers of oxidative stress. Expression of NOS2 was determined by immunohistochemistry 

and Western blot analysis while nitrate/nitrite levels were measured via spectral analysis. Total 

histopathological scoring of cellular damage was significantly decreased in hepatic IR injury 

following ASX treatment. Electron microscopy of postischemic tissue demonstrated parenchymal 

cell damage, swelling of mitochondria, disarrangement of rough endoplasmatic reticulum which 

was also partially reduced by ASX treatment. Astaxanthine treatment significantly decreased 

hepatic conversion of XDH to XO and tissue protein carbonyl levels following IR injury. The 

current results suggest that the mechanisms of action by which ASX reduces IR damage may 

include antioxidant protection against oxidative injury. 2009 Elsevier Ireland Ltd. All rights 

reserved. 

PMID: 19900500 [PubMed - indexed for MEDLINE 


305

Xenobiotica. 1996 Jan;26(1):49-63. 

Effects of canthaxanthin, astaxanthin, lycopene and lutein on liver 

xenobiotic-metabolizing enzymes in the rat. 

Gradelet S, Astorg P, Leclerc J, Chevalier J, Vernevaut MF, Siess MH. 

Unité de Toxicologie Nutritionnelle, Institut National de la Recherche 

Agronomique, DIJON, France. 

1. The catalytic activities of several phase I and II xenobiotic-metabolizing enzymes and 

the immunochemical detection of P4501A and 2B have been investigated in liver 

microsomes and cytosol of male rats fed for 15 days with diets containing canthaxanthin, 

astaxanthin, lycopene or lutein (as lutein esters) (300 mg/kg diet) and in rats fed 

increasing levels (10, 30, 100 and 300 ppm) of canthaxanthin or astaxanthin in the diet. 2. 

Canthaxanthin increased the liver content of P450, the activities of NADH- and NADPHcytochrome 

c reductase, and produced a substantial increase of some P450-dependent 

activities, especially ethoxyresorufin O-deethylase (EROD) (x 139) and 

methoxyresorufin O-demethylase (MROD) (x 26). Canthaxanthin also increased 

pentoxy-(PROD) and benzoxyresorufin O-dealkylases (BROD), but did not affect. 

NADPH-cytochrome c reductase and erythromycin N-demethylase (ERDM) activities 

and decreased nitrosodimethylamine N-demethylase (NDMAD) activity. Phase II p- 

nitrophenol UDP-glucuronosyl transferase (4NP-UGT) and quinone reductase (QR) 

activities were also increased by canthaxanthin treatment. These enhancing effects on 

EROD, MROD and 4NP-UGT were clearly detectable at a dose as low as 10 ppm of 

canthaxanthin in the diet; the induction of QR was only observed in rats fed > or = 100 

ppm. Astaxanthin induced the same pattern of enzymes activities as canthaxanthin, but to 

a lesser extent: its effects on phase I enzymes and 4NP-UGT were observed in rats fed > 

or = 100 ppm, and QR was not increased. Western blots of microsomal proteins clearly 

showed the induction of P4501A1 and 1A2 by canthaxanthin and astaxanthin. By 

contrast, lutein had no effect on the phase I and II xenobiotic-metabolizing enzymes 

activities measured. Lycopene only decreased NDMAD activity. 3. The two 4- 

oxocarotenoids canthaxanthin and astaxanthin are substantial inducers of liver P4501A1 

and 1A2 in the rat, and coinduce 4NP-UGT and QR, just like polycyclic aromatic 

hydrocarbon, beta-naphtoflavone or dioxin (TCDD). However, these latter classical 

P4501A inducers also induce aldehyde dehydrogenase class 3 (ALDH3); this enzyme is 

not increased, or only marginally, by canthaxanthin and astaxanthin. These two 

oxocarotenoids form a new class of inducers of P4501A, are structurally very different 

from the classical inducers quoted above, which are ligands of the AH receptor. 


 

 

In Vitro 




306

Food Chem Toxicol. 2008 Jan;46(1):212-9. Epub 2007 Aug 14. 

Effect of astaxanthin on kidney function impairment and oxidative 

stress induced by mercuric chloride in rats. 

Augusti PR, Conterato GM, Somacal S, Sobieski R, Spohr PR, Torres JV, 

Charão MF, Moro AM, Rocha MP, Garcia SC, Emanuelli T. 

Post-graduate Program on Toxicological Biochemistry, Center of Natural and 

Exact Sciences, Federal University of Santa Maria, 97105-900 Santa Maria, RS, 

Brazil. 

Reactive oxygen species are implicated as mediators of tissue damage in the acute 

renal failure induced by inorganic mercury. Astaxanthin (ASX), a carotenoid with 

potent antioxidant properties, exists naturally in various plants, algae, and 

seafoods. This paper evaluated the ability of ASX to prevent HgCl(2) 

nephrotoxicity. Rats were injected with HgCl(2) (0 or 5 mg/kg b.w., sc) 6h after 

ASX had been administered (0, 10, 25, or 50mg/kg, by gavage) and were killed 

12h after HgCl(2) exposure. Although ASX prevented the increase of lipid and 

protein oxidation and attenuated histopathological changes caused by HgCl(2) in 

kidney, it did not prevent creatinine increase in plasma and delta-aminolevulinic 

acid dehydratase inhibition induced by HgCl(2). Glutathione peroxidase and 

catalase activities were enhanced, while superoxide dismutase activity was 

depressed in HgCl(2)-treated rats when compared to control and these effects 

were prevented by ASX. Our results indicate that ASX could have a beneficial 

role against HgCl(2) toxicity by preventing lipid and protein oxidation, changes in 

the activity of antioxidant enzymes and histopathological changes. 



Additional Areas of Research: Renal Protective (Kidney) 

307

J Herb Pharmacother. 2005;5(1):17-26. 

A preliminary investigation of the enzymatic inhibition of 5alphareduction 

and growth of prostatic carcinoma cell line LNCap-FGC 

by natural astaxanthin and Saw Palmetto lipid extract in vitro. 

Anderson ML. 

Research and Development, Triarco Industries, Wayne, NJ 07470, USA. 

mark.anderson@triarco.com 

Inhibition of 5alpha-reductase has been reported to decrease the symptoms of 

benign prostate hyperplasia (BPH) and possibly inhibit or help treat prostate 

cancer. Saw Palmetto berry lipid extract (SPLE) is reported to inhibit 5alphareductase 

and decrease the clinical symptoms of BPH. Epidemiologic studies 

report that carotenoids such as lycopene may inhibit prostate cancer. In this 

investigation the effect of the carotenoid astaxanthin, and SPLE were examined 

for their effect on 5alpha-reductase inhibition as well as the growth of prostatic 

carcinoma cells in vitro. These studies support patent #6,277,417 B1. The results 

show astaxanthin demonstrated 98% inhibition of 5alpha-reductase at 300 

microg/mL in vitro. Alphastat, the combination of astaxanthin and SPLE, showed 

a 20% greater inhibition of 5alpha-reductase than SPLE alone n vitro. A nine day 

treatment of prostatic carcinoma cells with astaxanthin in vitro produced a 24% 

decrease in growth at 0.1 mcg/mL and a 38% decrease at 0.01 mcg/mL. SPLE 

showed a 34% decrease at 0.1 mcg/mL. CONCLUSIONS: Low levels of 

carotenoid astaxanthin inhibit 5alpha-reductase and decrease the growth of human 

prostatic cancer cells in vitro. Astaxanthin added to SPLE shows greater 

inhibition of 5alpha-reductase than SPLE alone in vitro. 


 

In Vitro 


Additional Areas of Research: Prostate Health 

308

Reprod Biomed Online. 2003 Oct-Nov;7(4):385-91. 

The role of food supplements in the treatment of the infertile man. 

Comhaire FH, Mahmoud A. 

Centre for Medical and Urological Andrology, Ghent University Hospital, De 

Pintelaan, 185, B 9000 Gent, Belgium. frank.comhaire@rug.ac.be 

Recently, concerns have been raised about the presumptive increased risk of 

serious undesirable side effects in children born after IVF and intracytoplasmic 

sperm injection (ICSI). These treatments must, therefore, be reserved as the 

ultimate option after evidence-based and cause-directed treatment of the male 

patient with deficient semen has been exhausted. The present authors found that 

sperm quality and function improved with the intake of complementary food 

supplementation using a combination of zinc and folic acid, or the antioxidant 

astaxanthin (Astacarox), or an energy-providing combination containing (actyl)- 

carnitine (Proxeed). Also, double blind trials showed that the latter two substances 

increase spontaneous or intrauterine insemination- (IUI-) assisted conception 

rates. Extracts of Pinus maritima bark (Pycnogenol), which inhibits the cyclooxygenase 

enzyme, reducing prostaglandin production and inflammatory reaction, 

and extracts of the Peruvian plant Lepidium meyenii were shown to improve 

sperm morphology and concentration, respectively, in uncontrolled trials. Linseed 

(flaxseed) oil contains alfa-linolenic acid and lignans. The former corrects the 

deficient intake of omega-3 essential fatty acids, which is correlated with 

impaired sperm motility among subfertile men. Lignans are precursors of 

enterolacton, which inhibits aromatase and reduces the ratio of 16-OH over 2-OH 

oestrogen metabolites. The resulting reduction in oestrogen load may favourably 

influence Sertoli cell function. 


Additional Areas of Research: Male Fertility 

309

Asian J Androl. 2005 Sep;7(3):257-62. 

Combined conventional/antioxidant "Astaxanthin" treatment for 

male infertility: a double blind, randomized trial. 

Comhaire FH, El Garem Y, Mahmoud A, Eertmans F, Schoonjans F. 

Ghent University Hospital, Department of Medical and Urological Andrology, 

9k12 IE, De Pintelaan, 185, B 9000, Gent, Belgium. frank.comhaire@ugent.be 

AIM: To evaluate the treatment of male infertility with a strong natural 

antioxidant, in addition to conventional treatment. METHODS: Using a double 

blind, randomized trial design, 30 men with infertility of > or =2 months and 

female partners with no demonstrable cause of infertility received conventional 

treatment according to the guidelines of the World Health Organization (WHO), 

and either a strong antioxidant Astaxanthin 16 mg/day (AstaCarox, AstaReal AB, 

Gustavsberg, Sweden) or placebo for 3 months. The effects of treatment on semen 

parameters, reactive oxygen species (ROS), zona-free hamster oocyte test, serum 

hormones including testosterone, luteinizing hormone (LH), follicle stimulating 

hormone (FSH) and Inhibin B, and spontaneous or intrauterine insemination 

(IUI)-induced pregnancies were evaluated. RESULTS: ROS and Inhibin B 

decreased significantly and sperm linear velocity increased in the Astaxanthin 

group (n = 11), but not in the placebo group (n = 19). The results of the zona-free 

hamster oocyte test tended to improve in the Astaxanthin group in contrast with 

the placebo group, though not reaching statistical significance. The total and per 

cycle pregnancy rates among the placebo cases (10.5 % and 3.6 %) were lower 

compared with 54.5 % and 23.1 % respectively in the Astaxanthin group (P = 

0.028; P = 0.036). CONCLUSION: Although the present study suggests a 

positive effect of Astaxanthin on sperm parameters and fertility, the results need 

to be confirmed in a larger trial before recommending Astaxanthin for the 

complementary treatment of infertile men. 


 

 

 

Clinical Trial 

Randomized Controlled Trial 




310

Asian J Androl. 2005 Sep;7(3):257-62 

Semen and Fertility Abstract for Astaxathin 

Comhaire FH, El Garem Y, Mahmoud A, Eertmans F, Schoonjans F. 

To evaluate the treatment of male infertility with a strong natural antioxidant, in 

addition to conventional treatment. METHODS: Using a double blind, 

randomized trial design, 30 men with infertility of > or =2 months and female 

partners with no demonstrable cause of infertility received conventional treatment 

according to the guidelines of the World Health Organization (WHO), and either 

a strong antioxidant Astaxanthin 16 mg/day (AstaCarox, AstaReal AB, 

Gustavsberg, Sweden) or placebo for 3 months. The effects of treatment on semen 

parameters, reactive oxygen species (ROS), zona-free hamster oocyte test, serum 

hormones including testosterone, luteinizing hormone (LH), follicle stimulating 

hormone (FSH) and Inhibin B, and spontaneous or intrauterine insemination 

(IUI)-induced pregnancies were evaluated. RESULTS: ROS and Inhibin B 

decreased significantly and sperm linear velocity increased in the Astaxanthin 

group (n = 11), but not in the placebo group (n = 19). The results of the zona-free 

hamster oocyte test tended to improve in the Astaxanthin group in contrast with 

the placebo group, though not reaching statistical significance. The total and per 

cycle pregnancy rates among the placebo cases (10.5 % and 3.6 %) were lower 

compared with 54.5 % and 23.1 % respectively in the Astaxanthin group (P = 

0.028; P = 0.036). CONCLUSION: Although the present study suggests a 

positive effect of Astaxanthin on sperm parameters and fertility, the results need 

to be confirmed in a larger trial before recommending Astaxanthin for the 

complementary treatment of infertile men 


311

XIII International Carotenoid Symposiumm Hawaii January 2002. Patent Cooperation 

Treaty Application 

WO99 / 29313. AstaCarotene AB, Sweden. 

Natural Astaxanthin Improves Semen Quality in Infertile Men 

GAREM, Y.E., A. LIGNELL u. F. COMHAIRE 

Natural Astaxanthin from Haematococcus Algae has been shown in a double 

blind, placebo controlled clinical to improve fertility in infertile men. Natural 

Astaxanthin had previously been shown to improve fertility in male animals such 

as boars and stallions. This study was conducted on men who were diagnosed as 

infertile due to abnormal sperm quality. The experimental group received 16 mg 

of Natural Astaxanthin per day for three months. The results were an 

improvement in conception rate in the experimental group by 478% over the 

placebo group. The scientist concluded that supplementation with Natural 

Astaxanthin improved the quality of the spermatozoa, which is suggested to be 

the plausible explanation for the increased frequency of conception.* 


312

Phytother Res. 2010 Jul 14. [Epub ahead of print] 

Summative interaction between astaxanthin, Ginkgo biloba extract 

(EGb761) and vitamin C in Suppression of respiratory inflammation: a 

comparison with ibuprofen. 

Haines DD, Varga B, Bak I, Juhasz B, Mahmoud FF, Kalantari H, Gesztelyi R, Lekli I, 

Czompa A, Tosaki A. 

Department of Pharmacology, Faculty of Pharmacy, University of Debrecen, Debrecen, 

Hungary. 


In this study, combinations of Ginkgo biloba leaf extract (EGb761) plus the carotenoid 

antioxidant astaxanthin (ASX) and vitamin C were evaluated for a summative dose effect 

in the inhibition of asthma-associated inflammation in asthmatic guinea-pigs. Ovalbuminsensitized 

Hartley guinea-pigs challenged with ovalbumin aerosol to induce asthma, were 

administered EGb761, ASX, vitamin C or ibuprofen. Following killing, bronchoalveolar 

lavage (BAL) fluid was evaluated for inflammatory cell infiltrates and lung tissue cyclic 

nucleotide content. Each parameter measured was significantly altered to a greater degree 

by drug combinations, than by each component acting independently. An optimal 

combination was identified that included astaxanthin (10 mg/kg), vitamin C (200 mg/kg) 

and EGb761 (10 mg/kg), resulting in counts of eosinophils and neutrophils each 1.6-fold 

lower; macrophages 1.8-fold lower, cAMP 1.4-fold higher; and cGMP 2.04-fold higher 

than levels in untreated, asthmatic animals (p < 0.05). In conclusion, EGb761, ASX and 

vitamin C are shown here to interact summatively to suppress inflammation with efficacy 

equal to or better than ibuprofen, a widely used non-steroidal antiinflammatory drug 

(NSAID). Such combinations of non-toxic phytochemicals constitute powerful tools for 

the prevention of onset of acute and chronic inflammatory disease if consumed regularly 

by healthy individuals; and may also augment the effectiveness of therapy for those with 

established illness. Copyright (c) 2010 John Wiley & Sons, Ltd. 


Additional Areas of Research: Asthma 

313

J Pharmacol Sci. 2004 Feb;94(2):129-36. 

In vitro effects of astaxanthin combined with ginkgolide B on T 

lymphocyte activation in peripheral blood mononuclear cells from 

asthmatic subjects. 

Mahmoud FF, Haines DD, Abul HT, Abal AT, Onadeko BO, Wise JA. 

Department of Medical Laboratory Sciences, Faculty of Allied Health Sciences, 

Kuwait University, Sulaibikhat. fadia@hsc.kuniv.edu.kw 

This study was undertaken to identify novel approaches to pharmacological 

treatment of asthma. Here we hypothesize that the platelet-activating factor 

receptor antagonist ginkgolide B (GB) in combination with the antioxidant 

carotenoid astaxanthin (ASX) suppresses T cell activation comparably to two 

commonly-used antihistamines: cetirizine dihydrochloride (CTZ) and azelastine 

(AZE). Peripheral blood mononuclear cells from asthmatics, cultured 24 h with 

either 50 microg/ml phytohemaglutinin (PHA) or PHA plus selected dosages of 

each drug are analyzed by flow cytometry for CD25+ or HLA-DR+ on CD3+ (T 

cells). Results are reported as stimulation indices (SI) of %CD3+CD25+ cells or 

%CD3+HLA-DR+ cells in cultures treated with PHA alone versus these 

subpopulations in cultures treated with both PHA and drugs. Combinations of 

ASX and GB exhibited optimal suppression at 10(-7) M GB + 10(-8) M ASX for 

CD3+CD25+ (SI = 0.79 +/- 0.04, P = 0.001) and 10(-7) M GB + 10(-7) M ASX 

for CD3+HLA-DR+ (SI = 0.82 +/- 0.05, P = 0.004). In conclusion, suppression of 

T cell activation below fully stimulated values by GB, ASX, and their 

combinations was comparable and for some combinations better than that 

mediated by CTZ and AZE. These results suggest that ASX and GB may have 

application as novel antiasthmatic formulations. 


 



Additional Areas of Research: Asthma 

314

Eur J Nutr. 2010 Apr 2. [Epub ahead of print] 

Astaxanthin addition improves human neutrophils function: in vitro study. 

Macedo RC, Bolin AP, Marin DP, Otton R. 

Postgraduate Program, Health Science, CBS, Cruzeiro do Sul University, Avenida 

Regente Feijó, 1295. Tatuapé, São Paulo, SP, CEP 03342-000, Brazil. 


PURPOSE: The aim of the present study was to evaluate the in vitro effect of carotenoid 

astaxanthin (ASTA) on the phagocytic and microbicidal capacities, cytokine release, and 

reactive oxygen species production in human neutrophils. 

METHODS: The following parameters were evaluated: cytotoxic effect of ASTA on 

human neutrophils viability, phagocytic and microbicidal capacities of neutrophils by 

using Candida albicans assay, intracellular calcium mobilization (Fura 2-AM fluorescent 

probe), superoxide anion (lucigenin and DHE probes), hydrogen peroxide (H(2)O(2), 

phenol red), and nitric oxide (NO.) (Griess reagent) production, activities of antioxidant 

enzymes (total/Mn-SOD, CAT, GPx, and GR), oxidative damages in biomolecules 

(TBARS assay and carbonyl groups), and cytokine (IL-6 and TNF-alpha) release. 

RESULTS: Astaxanthin significantly improves neutrophil phagocytic and microbicidal 

capacity, and increases the intracellular calcium concentration and NO. production. Both 

functional parameters were accompanied by a decrease in superoxide anion and hydrogen 

peroxide and IL-6 and TNF-alpha production. Oxidative damages in lipids and proteins 

were significantly decreased after ASTA-treatment. 

CONCLUSIONS: Taken together our results are supportive to a beneficial effect of 

astaxanthin-treatment on human neutrophils function as demonstrated by increased 

phagocytic and fungicide capacity as well as by the reduced superoxide anion and 

hydrogen peroxide production, however, without affecting neutrophils capacity to kill C. 

albicans. This process appears to be mediated by calcium released from intracellular 

storages as well as nitric oxide production. 


Additional Areas of Research: Candida 

315

Phytother Res. 2010 Jan;24(1):54-9. 

Cytoprotective role of astaxanthin against glycated protein/iron chelateinduced 

toxicity in human umbilical vein endothelial cells. 

Nishigaki I, Rajendran P, Venugopal R, Ekambaram G, Sakthisekaran D, Nishigaki Y. 

NPO International Laboratory of Biochemistry, 1-166 Uchide, Nakagawa-ku Nagoya 

454-0926, Japan. nishigaki@se.starcat.ne.jp 






interfering with ROS generation in these cells. Glycated fetal bovine serum (GFBS) was 

prepared by incubating fetal bovine serum (FBS) with high-concentration glucose. 

Stimulation of cultured HUVECs with 50 mm 1 mL of GFBS significantly enhanced 

lipid peroxidation and decreased antioxidant enzyme activities and levels of phase II 

enzymes. However, preincubation of the cultures with ASX resulted in a marked decrease 

in the level of lipid peroxide (LPO) and an increase in the levels of antioxidant enzymes 

in an ASX concentration-dependent manner. These results demonstrate that ASX could 

inhibit LPO formation and enhance the antioxidant enzyme status in GFBS/iron chelateexposed 

endothelial cells by suppressing ROS generation, thereby limiting the effects of 

the AGE-RAGE interaction. The results indicate that ASX could have a beneficial role 

against glycated protein/iron chelate-induced toxicity by preventing lipid and protein 

oxidation and increasing the activity of antioxidant enzymes. 


Additional Areas of Research: Toxicity 

316

Food Chem Toxicol. 2010 Jun;48(6):1741-5. Epub 2010 Apr 9. 

Astaxanthin improves the proliferative capacity as well as the osteogenic 

and adipogenic differentiation potential in neural stem cells. 

Kim JH, Nam SW, Kim BW, Kim WJ, Choi YH. 

Department of Biomaterial Control, Dong-Eui University, Busan, South Korea. 


In the present study, the effect of astaxanthin on improvement of the proliferative 

capacity as well as the osteogenic and adipogenic differentiation potential in neural stem 

cells (NSCs) was evaluated. Treatment of astaxanthin-induced actives cell growth in a 

dose-dependent and time-dependent manner. Results from a clonogenic assay clearly 

indicated that astaxanthin can actively stimulate proliferation of NSCs. Astaxanthininduced 

improvement in the proliferative capacity of NSCs resulted in overexpression of 

several proliferation-related proteins. Astaxanthin-induced activation of PI3K and its 

downstream mediators, p-MEK, p-ERK, and p-Stat3 in NSCs resulted in subsequent 

induction of expression of proliferation-related transcription factors (Rex1, CDK1, and 

CDK2) and stemness genes (OCT4, SOX2, Nanog, and KLF4). Astaxanthin also 

improved the osteogenic and adipogenic differentiation potential of NSCs. Astaxanthintreated 

NSCs showed prominent calcium deposits and fat formation. These results were 

consistent with overexpression of osteogenesis-related genes (osteonectin, RXR, and 

osteopontin) and adipogenesis-related genes (AP and PPAR-gamma) after astaxanthin 

treatment. These findings clearly demonstrated that astaxanthin acts synergistically on the 

regulatory circuitry that controls proliferation and differentiation of NSCs. Copyright 

2010 Elsevier Ltd. All rights reserved. 


Additional Areas of Research: Neural Stem Cells 

317

Eur J Pharm Sci. 2003 Jul;19(4):299-304. 

Oral bioavailability of the antioxidant astaxanthin in humans is 

enhanced by incorporation of lipid based formulations. 

Mercke Odeberg J, Lignell A, Pettersson A, Höglund P. 

Department of Clinical Pharmacology, Lund University Hospital, S-221 85 Lund, 

Sweden. johanna.odeberg@klinfarm.lu.se 

Astaxanthin is a carotenoid with antioxidant properties, synthesised by plants and 

algae, and distributed in marine seafood. Astaxanthin is also available as a food 

supplement, but, like other carotenoids, is a very lipophilic compound and has 

low oral bioavailability. However, bioavailability can be enhanced in the presence 

of fat. There is not much information in the literature about the pharmacokinetics 

of oral astaxanthin in humans. In this open parallel study, healthy male volunteers 

received a single dose of 40 mg astaxanthin, as lipid based formulations or as a 

commercially available food supplement, followed by blood sampling for further 

analysis of plasma concentrations. Pharmacokinetic parameters were calculated to 

evaluate the extent and rate of absorption from each formulation. The elimination 

half-life was 15.9+/-5.3 h (n=32), and showed a mono-phasic curve. Three lipid 

based formulations: long-chain triglyceride (palm oil) and polysorbate 80 

(formulation A), glycerol mono- and dioleate and polysorbate 80 (formulation B), 

and glycerol mono- and dioleate, polysorbate 80 and sorbitan monooleate 

(formulation C), all showed enhanced bioavailability, ranging from 1.7 to 3.7 

times that of the reference formulation. The highest bioavailability was observed 

with formulation B, containing a high content of the hydrophilic synthetic 

surfactant polysorbate 80. 


 



Additional Areas of Research: Bioavailability 

318

AKVAFORSK (Inst. Aquaculture Research AS), N-6600 Sunndalsøra, Norway 

On bioavailability and deposition of bent Z-isomers of astaxanthin 

Marianne Østerlie, Bjørn Bjerkeng* and Synnøve Liaaen-Jensen 

Experiments have been performed in which rainbow trout (Oncorhynchus mykiss) 

were fed diets containing a mixture of the all-E, 9Z, and 13Z geometrical isomers 

of astaxanthin or three male middle-aged human subjects were administered a 

single dose containing a similar astaxanthin isomer mixture. In rainbow trout, 

selective accumulation of all-E-astaxanthin was observed in tissues and blood 

plasma, and of 13Z-astaxanthin in the liver. In human blood plasma, 13Zastaxanthin 

appeared to accumulate, and the distribution of the astaxanthin E/Z 

isomers remained constant in the mixed chylomicron and very low density 

(VLDL), and low density (LDL) and high density (HDL) lipoprotein fractions. In 

conclusion, more attention than assumed in the past must be paid to the E/Z 

configuration of xanthophylls when bioavailability and functional aspects are 

concerned in different species. 


319

J Nutr Biochem. 2000 Oct;11(10):482-90. 

Plasma appearance and distribution of astaxanthin E/Z and R/S 

isomers in plasma lipoproteins of men after single dose 

administration of astaxanthin. 

Østerlie M, Bjerkeng B, Liaaen-Jensen S. 

HIST, Department of Food Science, N-7004, Trondheim, Norway. 

Appearance, pharmacokinetics, and distribution of astaxanthin E/Z and R/S 

isomers in plasma and lipoprotein fractions were studied in 3 middle-aged male 

volunteers (37-43 years) after ingestion of a single meal containing a 100 mg dose 

of astaxanthin. The astaxanthin source consisted of 74% all-E-, 9% 9Z-, 17% 

13Z-astaxanthin (3R,3'R-, 3R,3'S; meso-, and 3S,3'S-astaxanthin in a 1:2:1 ratio). 

The plasma astaxanthin concentration--time curves were measured during 72 hr. 

Maximum levels of astaxanthin (1.3 +/- 0.1 mg/L) were reached 6.7 +/- 1.2 hr 

after administration, and the plasma astaxanthin elimination half-life was 21 +/- 

11 hr. 13Z-Astaxanthin accumulated selectively, whereas the 3 and 3'R/S 

astaxanthin distribution was similar to that of the experimental meal. Astaxanthin 

was present mainly in very low-density lipoproteins containing chylomicrons 

(VLDL/CM; 36-64% of total astaxanthin), whereas low-density lipoprotein 

(LDL) and high-density lipoprotein (HDL) contained 29% and 24% of total 

astaxanthin, respectively. The astaxanthin isomer distribution in plasma, 

VLDL/CM, LDL, and HDL was not affected by time. The results indicate that a 

selective process increases the relative proportion of astaxanthin Z-isomers 

compared to the all-E-astaxanthin during blood uptake and that astaxanthin E/Z 

isomers have similar pharmacokinetics. 



320

Oxid Med Cell Longev. 2011;2011:596240. Epub 2011 Oct 12. 

Supplemental Cellular Protection by a 

Carotenoid Extends Lifespan via Ins/IGF- 

1 Signaling in Caenorhabditis elegans. 

Yazaki K, Yoshikoshi C, Oshiro S, Yanase S. 

Source 

Department of Health Science, Daito Bunka University School of Sports and Health 

Science, Iwadono 560, Higashi-matsuyama, Saitama 355-8501, Japan. 


Astaxanthin (AX), which is produced by some marine animals, is a type of carotenoid 

that has antioxidative properties. In this study, we initially examined the effects of AX on 

the aging of a model organism C. elegans that has the conserved intracellular pathways 

related to mammalian longevity. The continuous treatments with AX (0.1 to 1 mM) from 

both the prereproductive and young adult stages extended the mean lifespans by about 

16-30% in the wild-type and long-lived mutant age-1 of C. elegans. In contrast, the AXdependent 

lifespan extension was not observed even in a daf-16 null mutant. Especially, 

the expression of genes encoding superoxide dismutases and catalases increased in two 

weeks after hatching, and the DAF-16 protein was translocated to the nucleus in the AXexposed 

wild type. These results suggest that AX protects the cell organelle mitochondria 

and nucleus of the nematode, resulting in a lifespan extension via an Ins/IGF-1 signaling 

pathway during normal aging, at least in part. 

PMID: 22013497 [PubMed - in process] 


Additional Areas of Astaxanthin Research: Anti-Aging 

321

Editors’ Note: A complete Safety Profile for BioAstin® Natural 

Astaxanthin, a trademarked Astaxanthin product from Haematococcus 

Pluvialis microalgae is available at www.cyanotech.com 

Food Chem Toxicol. 2008 Sep;46(9):3030-6. Epub 2008 Jun 10. 

Safety assessment of astaxanthin-rich microalgae biomass: Acute 

and subchronic toxicity studies in rats. 

Stewart JS, Lignell A, Pettersson A, Elfving E, Soni MG. 

Covance Laboratories Ltd., Otley Road, Harrogate, North Yorkshire, HG3 1PY 

England, United Kingdom. 

Astaxanthin, a natural nutritional component, is marketed as a dietary supplement 

around the world. The primary commercial source for astaxanthin is 

Haematococcus pluvialis (microalgae). The objective of the present study was to 

investigate the acute and subchronic toxicity of an astaxanthin-rich biomass of H. 

pluvialis (AstaCarox). The oral LD(50) of the biomass in rats was greater than 

12g/kg body weight. In the subchronic study, Wistar rats (10/sex/group) were fed 

diets containing 0%, 1%, 5% and 20% of the biomass (weight/weight) for 90 

days. trans-Astaxanthin was quantifiable in the plasma of the high-dose treated 

group only. Compared to the control group, no treatment-related biologically 

significant effects of astaxanthin were noted on body weight or body weight gain. 

Biomass feeding did not affect hematological parameters. In the high-dose group, 

slightly elevated alkaline phosphatase and changes in some urine parameters and 

an increase in kidney weight in both sexes were noted. Histopathology 

examinations did not reveal adverse effects except for a marginal increase in 

pigment in the straight proximal tubule of the kidney in 5/10 female rats treated 

with the high-dose. These changes were not considered as toxicologically 

significant. Although the rats in high-dose group received about 9% more fat, it is 

unlikely that this confounding factor significantly altered the outcome. The noobserved 

adverse-effect-levels (NOAEL) of the astaxanthin-rich biomass for male 

and female rats were determined as 14,161 and 17,076mg/kg body weight/day, or 

465 and 557mg astaxanthin/kg/day, respectively, the highest dose tested. 



Additional Areas of Research: Safety 

322

Astaxanthin Abstract Book 2013 - Cyanotech

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